Genomic Variability in the Neuraminidase Gene and Antiviral Resistance of Influenza A(H1N1)pdm09 Virus Isolated from Acute Respiratory Infection Patients in Shiraz, Iran (2023–2024)
preprint
OA: closed
CC-BY-4.0
Abstract
Abstract Background The 2009 Influenza A(H1N1)pdm09 pandemic caused significant global morbidity and mortality. Mutations in the neuraminidase (NA) gene can influence viral pathogenicity, immune evasion, and resistance to antiviral drugs, necessitating continuous molecular surveillance to guide antiviral development. Methods From November 2023 to December 2024, 300 respiratory samples were collected from patients with acute respiratory infections in Shiraz, Iran. Real-Time PCR identified Influenza A(H1N1)pdm09, followed by conventional PCR amplification and sequencing of the NA gene. Mutations and resistance to neuraminidase inhibitors (oseltamivir, zanamivir) were assessed using MEGA 11, I-TASSER modeling, and AutoDock Vina docking. Phylogenetic analysis was performed to evaluate genetic relationships. Results Of the 300 samples, 14 were positive for Influenza A(H1N1)pdm09. Sequence analysis identified 26 NA gene mutations, with N44S, I106V, and N200S detected in 3, 4, and 3 isolates, respectively. No isolates showed the H275Y mutation, indicating sensitivity to oseltamivir and zanamivir. Phylogenetically, the isolates were highly similar to A/California/07/2009, clustering with 2008–2009 global strains from Luxembourg and Yamagata. Conclusion Despite significant genetic variability in the NA gene, current antiviral therapies remain effective against these isolates. This study underscores the importance of ongoing surveillance to detect emerging mutations that could impact influenza management globally.
Full text
107,890 characters
· extracted from
preprint-html
· click to expand
Genomic Variability in the Neuraminidase Gene and Antiviral Resistance of Influenza A(H1N1)pdm09 Virus Isolated from Acute Respiratory Infection Patients in Shiraz, Iran (2023–2024) | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Genomic Variability in the Neuraminidase Gene and Antiviral Resistance of Influenza A(H1N1)pdm09 Virus Isolated from Acute Respiratory Infection Patients in Shiraz, Iran (2023–2024) Mohammad Javad Shahbazi Nezhad, Neda Pirbonyeh, Mojtaba Mortazavi, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6495442/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background The 2009 Influenza A(H1N1)pdm09 pandemic caused significant global morbidity and mortality. Mutations in the neuraminidase (NA) gene can influence viral pathogenicity, immune evasion, and resistance to antiviral drugs, necessitating continuous molecular surveillance to guide antiviral development. Methods From November 2023 to December 2024, 300 respiratory samples were collected from patients with acute respiratory infections in Shiraz, Iran. Real-Time PCR identified Influenza A(H1N1)pdm09, followed by conventional PCR amplification and sequencing of the NA gene. Mutations and resistance to neuraminidase inhibitors (oseltamivir, zanamivir) were assessed using MEGA 11, I-TASSER modeling, and AutoDock Vina docking. Phylogenetic analysis was performed to evaluate genetic relationships. Results Of the 300 samples, 14 were positive for Influenza A(H1N1)pdm09. Sequence analysis identified 26 NA gene mutations, with N44S, I106V, and N200S detected in 3, 4, and 3 isolates, respectively. No isolates showed the H275Y mutation, indicating sensitivity to oseltamivir and zanamivir. Phylogenetically, the isolates were highly similar to A/California/07/2009, clustering with 2008–2009 global strains from Luxembourg and Yamagata. Conclusion Despite significant genetic variability in the NA gene, current antiviral therapies remain effective against these isolates. This study underscores the importance of ongoing surveillance to detect emerging mutations that could impact influenza management globally. Biological sciences/Biotechnology Biological sciences/Microbiology Neuraminidase Influenza A(H1N1)pdm09 oseltamivir resistance mutation phylogenetic analysis molecular surveillance Figures Figure 1 Figure 2 Introduction Influenza viruses, members of the Orthomyxoviridae family, are significant global health threats, causing seasonal epidemics and occasional pandemics( 1 ). Annually, influenza affects 5 to 15% of the population, leading to 3 to 5 million severe cases and up to 650,000 respiratory-related deaths worldwide( 2 ). Influenza A viruses, particularly the A(H1N1)pdm09 strain that emerged in 2009, are of major concern due to their high morbidity, mortality, and economic impact( 3 ). Shiraz, a major travel hub in Iran, experiences high influenza transmission, yet molecular data are scarce( 4 ). These single-stranded, negative-sense RNA viruses possess a segmented genome of eight distinct segments encoding essential proteins, including the surface glycoproteins hemagglutinin (HA) and neuraminidase (NA)( 5 – 8 ). The NA protein, encoded by the sixth genomic segment, facilitates viral release from infected cells by cleaving sialic acid residues, enabling spread within the respiratory tract( 9 , 10 ). NA also contributes to viral tropism through its specificity for α-2,6- and α-2,3-linked sialic acids on host cells( 11 ). However, the NA gene is highly mutable, undergoing antigenic drift (point mutations) and antigenic shift (genetic reassortment), which drive the emergence of new strains capable of evading immunity and developing antiviral resistance( 12 , 13 ). Neuraminidase inhibitors (NAIs), such as oseltamivir and zanamivir, are critical for influenza treatment and prophylaxis by blocking NA activity, but resistance—often linked to mutations like H275Y—poses a growing challenge( 14 – 16 ). Influenza A(H1N1)pdm09, responsible for the 2009 pandemic, continues to circulate and evolve, necessitating ongoing molecular surveillance( 3 , 17 ). Mutations in the NA gene can alter pathogenicity, immune evasion, and drug susceptibility, making their characterization essential for effective influenza management( 18 ). This study investigates the genomic changes in the NA gene of A(H1N1)pdm09 isolates from patients with acute respiratory infections in Shiraz, Iran, during 2024–2025, focusing on antiviral resistance and phylogenetic relationships to inform global surveillance efforts. Methods Sample Collection Between November 2023 and December 2024, 300 respiratory samples (nasopharyngeal swabs and sputum) were collected from patients at Shiraz teaching hospitals presenting with acute respiratory infection symptoms, including fever, cough, myalgia, and sore throat. Patients with confirmed co-infections or other respiratory viral infections, identified via multiplex PCR for common respiratory pathogens (e.g., RSV, SARS-CoV-2), were excluded. Samples were immediately placed in viral transport media and stored at -80°C until analysis. Ethical Approval This study was approved by the Ethics Committee of Shiraz University of Medical Sciences, Shiraz, Iran (approval number: IR.SUMS.MED.REC.1403.557). Informed consent was obtained from all patients or their legal guardians prior to sample collection. All procedures adhered to the Declaration of Helsinki, ensuring participant confidentiality and anonymity throughout the study. Data Availability The datasets generated and/or analyzed during the current study are available in the GenBank repository, under the accession numbers PQ836530 to PQ836543 . These sequences can be accessed through the following link: https://www.ncbi.nlm.nih.gov/nuccore/PQ836530-PQ836543. RNA Extraction and Real-Time RT-PCR for Detection of Influenza A(H1N1)pdm09 Viral RNA was extracted from the 300 samples using the High Pure Viral RNA Kit (Roche, Germany) following the manufacturer’s protocol, with extracted RNA stored at -80°C to prevent degradation. Real-time RT-PCR was performed using the SuperScript III Platinum One-Step Quantitative RT-PCR Kit (Invitrogen, USA) on a Rotor-Gene 6000 system (Corbett). The reaction mixture (20 μL) consisted of 4 μL RNA, 10 μL 2X master mix, 0.4 μL each of forward and reverse primers (10 μM), 0.4 μL probe (5 μM), and 4.8 μL RNase/DNase-free water. Primers and probes were designed according to the CDC 2024 protocol for Influenza A(H1N1)pdm09 detection (sequences available at [CDC reference or state ‘proprietary’ if not public]). Thermal cycling conditions were: reverse transcription at 50°C for 30 minutes, enzyme inactivation at 95°C for 2 minutes, followed by 45 cycles of 95°C for 15 seconds, 55°C for 30 seconds, and 72°C for 30 seconds. Of the 300 samples, 14 tested positive for Influenza A(H1N1)pdm09. Amplification and Sequencing of the Neuraminidase Gene RNA from the 14 positive samples was reverse-transcribed to cDNA using the SuperScript III First-Strand Synthesis System (Invitrogen, USA). The full-length NA gene was amplified via conventional PCR using primers specific for the A(H1N1)pdm09 NA gene (forward: 5’-AGCAAAAGCAGGAGT-3’, reverse: 5’-AGTAGAAACAAGGAGT-3’, designed to amplify the 1410 bp NA coding region). PCR reactions (50 μL) contained 5 μL cDNA, 25 μL 2X PCR master mix (TaKaRa, Japan), 1 μL each primer (10 μM), and 18 μL nuclease-free water. Cycling conditions were: 95°C for 3 minutes, followed by 35 cycles of 95°C for 30 seconds, 55°C for 30 seconds, and 72°C for 90 seconds, with a final extension at 72°C for 5 minutes. Amplicons were verified on a 2% agarose gel, purified using the QIAquick PCR Purification Kit (Qiagen, Germany), and sequenced by TopazGene (Iran) using Sanger sequencing. Phylogenetic and Mutation Analysis NA sequences were aligned with the reference strain A/California/07/2009 (GenBank: NC_026434.1) using ClustalW in MEGA 11 software. Phylogenetic trees were constructed via the maximum-likelihood method with the Tamura-Nei model, supported by 1000 bootstrap replicates. The 14 NA sequences were deposited in GenBank under accession numbers PQ836530 to PQ836543. Amino acid substitutions were identified by comparing the sequences to A/California/07/2009, focusing on positions associated with antiviral resistance, particularly H275Y for oseltamivir resistance. Structural Modeling and Antiviral Resistance Assessment The NA protein structure was modeled using I-TASSER (C-score: -2.05, TM-score: 0.47±0.15). Physicochemical properties were calculated with ProtParam (ExPASy). Molecular docking was performed using AutoDock Vina (v1.1.2) to assess the impact of mutations on inhibitor binding. Ligands (oseltamivir, zanamivir, peramivir, laninamivir) were retrieved from PubChem, converted to PDBQT format using Open Babel (v2.3.1), and docked against the NA model with a grid box centered on the active site (size: 30×30×30 Å, exhaustiveness: 8). Binding poses were visualized in PyMOL (v2.5). Results Detection of Influenza A(H1N1)pdm09 in Respiratory Samples From 300 throat and nasal swab samples collected between November 2023 and December 2024, 14 tested positive for Influenza A(H1N1)pdm09 via Real-Time PCR, confirmed by RT-PCR. The patients, aged 35 to 85 years, included 9 males and 5 females (Table 1), all of whom required hospitalization due to severe respiratory symptoms, including fever, cough, myalgia, and sore throat. Molecular Characterization of Neuraminidase Gene The NA sequences of the 14 A(H1N1)pdm09 isolates were submitted to GenBank under accession numbers PQ836530 to PQ836543 (Table 1). Phylogenetic analysis revealed that these isolates share high nucleotide similarity with the reference strain A/California/07/2009, clustering closely with 2008–2009 global strains from Luxembourg and Yamagata (Fig. 1). Amino Acid Variations in Neuraminidase Sequence analysis identified 26 amino acid substitutions in the NA gene compared to A/California/07/2009, with N44S, I106V, and N200S being the most frequent, observed in 3, 4, and 3 isolates, respectively (Table 2). Other mutations included A20T, I34V, Add36H, L40I, V81I, S90P, R107A, K111R, K143E, I241V, F256L, V264I, N270K, F306I, I314M, I321V, K369N, N386K, N386S, I389T, V394I, K432E, and D451G (Table 3). No isolates exhibited the H275Y mutation associated with oseltamivir resistance, indicating retained sensitivity to neuraminidase inhibitors oseltamivir and zanamivir. Structural Analysis of Neuraminidase Mutations The NA protein was modeled using I-TASSER (C-score: -2.05, TM-score: 0.47±0.15). Its physicochemical properties, calculated via ProtParam, showed a theoretical pI of 6.11, an instability index of 32.91, and an aliphatic index of 74.16, indicating moderate stability (Table 4). Molecular docking with inhibitors (peramivir, oseltamivir, zanamivir, and laninamivir) revealed that key mutations, including N44S, I34V, I106V, and others, are located distantly from the inhibitor-binding sites (Fig. 2), suggesting minimal impact on drug efficacy. Superimposition of the I-TASSER model with the 2hu4 crystal structure confirmed structural conservation in the catalytic head domain (Supplementary Fig. 1) Discussion Influenza A(H1N1)pdm09 remains a significant public health concern due to its potential for rapid evolution and antiviral resistance, necessitating continuous molecular surveillance(19). This study identified 26 amino acid substitutions in the NA gene of 14 A(H1N1)pdm09 isolates from Shiraz, Iran, collected between November 2023 and December 2024. The most frequent mutations—N44S, I106V, and N200S—were detected in 3, 4, and 3 isolates, respectively, but did not confer resistance to oseltamivir or zanamivir, as no H275Y mutations were observed. This aligns with prior studies in Shiraz from 2012–2013, which reported no oseltamivir resistance, though a 2016 study identified two resistant isolates with the H275Y mutation(20). Globally, resistance patterns vary; for instance, 34.3% of isolates in Argentina (2008) and 1.4% in Mexico (2009–2010) exhibited oseltamivir resistance due to H275Y, highlighting the importance of regional surveillance(20, 21). Phylogenetic analysis revealed that our isolates are closely related to the reference strain A/California/07/2009, clustering with 2008–2009 global strains from Luxembourg and Yamagata. This conserved lineage suggests limited antigenic drift in the NA gene over the past 15 years, consistent with findings from Australia (2001–2005) where oseltamivir resistance emerged sporadically without widespread drift(22). However, the presence of mutations like N44S and I106V, which are distal from the NA active site as shown by molecular docking, indicates ongoing genetic variability that could influence future pathogenicity or immune evasion(23). Similar distal mutations were reported in Shiraz in 2016 (e.g., V241I, N369K), suggesting a recurring pattern of variation in this region(20). The absence of oseltamivir resistance in our isolates supports the continued efficacy of NA inhibitors for managing A(H1N1)pdm09 infections in Shiraz. However, the genetic diversity observed underscores the need for vigilance, as mutations near functional sites could eventually impact inhibitor binding, as seen with the S247N mutation reported in other studies(22). Computational modeling, despite limitations such as incomplete loop representation, provides a rapid tool for assessing mutation impacts, which is critical for timely antiviral development(24). Future studies should integrate experimental validation (e.g., enzymatic assays) to confirm the functional effects of these mutations and extend surveillance to other viral proteins like hemagglutinin to fully understand immune escape potential. Conclusion This study demonstrates significant genetic variability in the NA gene of Influenza A(H1N1)pdm09 isolates from Shiraz, Iran, with 26 mutations identified, though none conferred resistance to oseltamivir or zanamivir. The phylogenetic similarity to A/California/07/2009 highlights a conserved lineage, while the absence of the H275Y mutation supports the ongoing effectiveness of NA inhibitors in this region. These findings emphasize the importance of continuous molecular surveillance to detect emerging mutations that could affect influenza management globally, informing antiviral strategies and public health responses. Declarations Author Contribution AmirReza Radfar primarily contributed to the manuscript writing, including the design of the study, data analysis, and interpretation. Mojtaba Mortazavi provided expertise in biotechnology, specifically assisting with the bioinformatics analysis, including structural modeling and molecular docking studies. Mohammad Javad Shahbazi Nezhad, Neda Pirbonyeh, Jamal Sarvari, and Afagh Moattari were responsible for the virology aspects of the study, including sample collection, virological testing, and the molecular characterization of Influenza A(H1N1)pdm09 strains. Afagh Moattari also contributed significantly to the manuscript writing, particularly in the interpretation of the results and the revision of the manuscript. All authors contributed to the drafting and critical revision of the manuscript and approved the final version for submission. References Kumari R, Sharma SD, Kumar A, Ende Z, Mishina M, Wang Y, et al. Antiviral approaches against influenza virus. Clinical microbiology reviews. 2023;36(1):e00040-22. EDITION T. The global impact of respiratory. Mahmud S, Tamanna RK. Influenza Virus Types, Subtypes and Genomic Lineage with Its Prevention and Control: A Narrative Review. Journal of Monno Medical College. 2024;10(1):43-51. Faramarzi H, Mousavi-Roknabadi RS, Hemmati A, Faramarzi A, Bakhtiari H. Epidemiology of Influenza in Fars Province, Southern Iran; a Population-Based Study (2015–2019). Archives of Iranian Medicine. 2021;24(3):199-208. Noda T. Selective genome packaging mechanisms of influenza A viruses. Cold Spring Harbor Perspectives in Medicine. 2021;11(7):a038497. Flerlage T, Boyd DF, Meliopoulos V, Thomas PG, Schultz-Cherry S. Influenza virus and SARS-CoV-2: pathogenesis and host responses in the respiratory tract. Nature Reviews Microbiology. 2021;19(7):425-41. Russell CJ. Hemagglutinin stability and its impact on influenza A virus infectivity, pathogenicity, and transmissibility in avians, mice, swine, seals, ferrets, and humans. Viruses. 2021;13(5):746. Moattari A, Emami A, Moghadami M, Honarvar B. Influenza viral infections among the Iranian Hajj pilgrims returning to Shiraz, Fars province, Iran. Influenza and other respiratory viruses. 2012;6(6):e77-e9. Li Y, Liu D, Wang Y, Su W, Liu G, Dong W. The importance of glycans of viral and host proteins in enveloped virus infection. Frontiers in Immunology. 2021;12:638573. Chauhan RP, Gordon ML. An overview of influenza A virus genes, protein functions, and replication cycle highlighting important updates. Virus genes. 2022;58(4):255-69. Burzyńska P, Sobala ŁF, Mikołajczyk K, Jodłowska M, Jaśkiewicz E. Sialic acids as receptors for pathogens. Biomolecules. 2021;11(6):831. Perofsky AC, Huddleston J, Hansen CL, Barnes JR, Rowe T, Xu X, et al. Antigenic drift and subtype interference shape A (H3N2) epidemic dynamics in the United States. Elife. 2024;13:RP91849. Dapat C, Saito R, Kyaw Y, Myint YY, Oo HN, Oo KY, et al. Delayed emergence of oseltamivir-resistant seasonal influenza A (H1N1) and pandemic influenza A(H1N1)pdm09 viruses in Myanmar. Influenza Other Respir Viruses. 2013;7(5):766-71. Yang JR, Huang YP, Chang FY, Hsu LC, Huang HY, Pan YT, et al. Characterization of oseltamivir‐resistant influenza A (H1N1) pdm09 viruses in Taiwan in 2009–2011. Journal of Medical Virology. 2013;85(3):379-87. Russell RJ, Haire LF, Stevens DJ, Collins PJ, Lin YP, Blackburn GM, et al. The structure of H5N1 avian influenza neuraminidase suggests new opportunities for drug design. Nature. 2006;443(7107):45-9. Goka E, Vallely P, Mutton K, Klapper P. Mutations associated with severity of the pandemic influenza A (H1N1) pdm09 in humans: a systematic review and meta-analysis of epidemiological evidence. Archives of Virology. 2014;159:3167-83. Medina RA, García-Sastre A. Influenza A viruses: new research developments. Nature Reviews Microbiology. 2011;9(8):590-603. McAuley JL, Gilbertson BP, Trifkovic S, Brown LE, McKimm-Breschkin JL. Influenza virus neuraminidase structure and functions. Frontiers in microbiology. 2019;10:39. Wang X, Xiao F, Wang Y, Deng X, Chen Z, Dong X, et al. Changed epidemiology of narcolepsy before, during, and after the 2009 H1N1 pandemic: a nationwide narcolepsy surveillance network study in mainland China, 1990–2017. Sleep. 2023;46(3):zsac325. Dia N, Niang MN, Diadhiou SA, Goudiaby DG, Faye A, Kiori D, et al. Spread of Influenza A (H1N1) oseltamivir-resistant viruses in Africa in 2008 confirmed by multiple introductions in Senegal. BMC Infectious Diseases. 2013;13:1-7. Pontoriero A, Baumeister E, Campos AM, Savy VL. Vigilancia virologica y resistencia a los antiviricos del virus de la gripe humana en la Argentina, 2005-2008. Revista Panamericana de Salud Publica. 2011;30:634+. Tamura D, DeBiasi RL, Okomo-Adhiambo M, Mishin VP, Campbell AP, Loechelt B, et al. Emergence of Multidrug-Resistant Influenza A(H1N1)pdm09 Virus Variants in an Immunocompromised Child Treated With Oseltamivir and Zanamivir. The Journal of Infectious Diseases. 2015;212(8):1209-13. Nguyen TH, Anh Pham NQ, Thai QM, Vu VV, Ngo ST, Horng J-T. Discovering Influenza Virus Neuraminidase Inhibitors via Computational and Experimental Studies. ACS Omega. 2024;9(49):48505-11. Vavricka CJ, Liu Y, Kiyota H, Sriwilaijaroen N, Qi J, Tanaka K, et al. Influenza neuraminidase operates via a nucleophilic mechanism and can be targeted by covalent inhibitors. Nat Commun. 2013;4:1491. Tables Table 1 Demographic and Genetic Details of Influenza A(H1N1)pdm09 Strains Sex Gene Bank Accession Origin Influenza A(H1N1) pdm09 Strains Name F M M M F M F M M M F M M M PQ836530 PQ836531 PQ836532 PQ836533 PQ836534 PQ836535 PQ836536 PQ836537 PQ836538 PQ836539 PQ836540 PQ836541 PQ836542 PQ836543 Shiraz Shiraz Shiraz Shiraz Shiraz Shiraz Shiraz Shiraz Shiraz Shiraz Shiraz Shiraz Shiraz Shiraz Influenza A virus (A/Iran/01/2024(H1N1)) Influenza A virus (A/Iran/02/2024(H1N1)) Influenza A virus (A/Iran/03/2024(H1N1)) Influenza A virus (A/Iran/04/2024(H1N1)) Influenza A virus (A/Iran/05/2024(H1N1)) Influenza A virus (A/Iran/06/2024(H1N1)) Influenza A virus (A/Iran/07/2024(H1N1)) Influenza A virus (A/Iran/08/2024(H1N1)) Influenza A virus (A/Iran/09/2024(H1N1)) Influenza A virus (A/Iran/10/2024(H1N1)) Influenza A virus (A/Iran/11/2024(H1N1)) Influenza A virus (A/Iran/12/2024(H1N1)) Influenza A virus (A/Iran/13/2024(H1N1)) Influenza A virus (A/Iran/14/2024(H1N1)) Table 2 Amino Acid Variation Across Influenza A(H1N1)pdm09 Isolates Isolates Number of amino acid 20 34 36 40 44 81 90 106 107 111 143 200 241 256 264 270 306 314 321 369 386 386 389 394 432 451 A/California/07/2009 A/shiraz/01 S A/shiraz/02 V I S V R S I K M V K E A/shiraz/03 V N I A/shiraz/04 T H P I A/shiraz/05 I A/shiraz/06 H S G A/shiraz/07 H S V S I A/shiraz/08 A/shiraz/09 H A/shiraz/10 A A/shiraz/11 H S V E S A/shiraz/12 A/shiraz/13 S A/shiraz/14 H S V S L T Table 3 Mutations in Influenza A(H1N1)pdm09 Isolates by Gene Bank Accession Gene Bank Accession Mutation PQ836530 N44S PQ836531 I34V, L401, I106V, K111R PQ836532 I241V, K369N PQ836533 A20T, S90P, F306I PQ836534 V81I PQ836535 N386S PQ836536 V394I PQ836538 Add 36H PQ836539 R107A PQ836540 K143E, N200S PQ836542 N44S PQ836543 F256L, I389T Table 4 Biochemical and Structural Parameters of Neuraminidase in Influenza A(H1N1)pdm09 Isolates Parameter Value Theoretical pI 6.11 Molecular weight 51613.01 Sequence length 469 Extinction coefficients (M-1 cm-1 at 260 nm)* 109985 − 108860 Asp + Glu 40 Arg + Lys 37 Instability index 32.91 Grand average of hydropathicity -0.259 Aliphatic index 74.16 Additional Declarations No competing interests reported. Supplementary Files SupplementaryFigures.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6495442","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":448286777,"identity":"7f569bd0-aa54-41cf-a5c3-6df00516a545","order_by":0,"name":"Mohammad Javad Shahbazi Nezhad","email":"","orcid":"","institution":"Shiraz University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Mohammad","middleName":"Javad Shahbazi","lastName":"Nezhad","suffix":""},{"id":448286780,"identity":"2e1ad1a3-417a-457d-8941-0b6eb4813dbb","order_by":1,"name":"Neda Pirbonyeh","email":"","orcid":"","institution":"Shiraz University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Neda","middleName":"","lastName":"Pirbonyeh","suffix":""},{"id":448286783,"identity":"27b6ca8d-e98a-4442-88af-f99896603062","order_by":2,"name":"Mojtaba Mortazavi","email":"","orcid":"","institution":"Graduate University of Advanced Technology","correspondingAuthor":false,"prefix":"","firstName":"Mojtaba","middleName":"","lastName":"Mortazavi","suffix":""},{"id":448286784,"identity":"ffbb71b5-6aaf-4878-b4e2-960c6725bea6","order_by":3,"name":"AmirReza Radfar","email":"data:image/png;base64,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","orcid":"","institution":"Shiraz University of Medical Sciences","correspondingAuthor":true,"prefix":"","firstName":"AmirReza","middleName":"","lastName":"Radfar","suffix":""},{"id":448286787,"identity":"7beebb37-ea43-48ef-86bb-4939c269608c","order_by":4,"name":"Jamal Sarvari","email":"","orcid":"","institution":"Shiraz University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Jamal","middleName":"","lastName":"Sarvari","suffix":""},{"id":448286789,"identity":"4ea4e080-a17b-4451-b50e-64ccae2c9cba","order_by":5,"name":"Afagh Moattari","email":"","orcid":"","institution":"Shiraz University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Afagh","middleName":"","lastName":"Moattari","suffix":""}],"badges":[],"createdAt":"2025-04-21 11:08:14","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6495442/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6495442/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":81954150,"identity":"907abd99-e714-488f-a54c-0225cad10b7c","added_by":"auto","created_at":"2025-05-05 09:41:39","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":294620,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePhylogenetic Analysis of Neuraminidase Gene Sequences of Shiraz Influenza A(H1N1)pdm09 Strains\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6495442/v1/ea4f5ffca9c31b66bee41178.png"},{"id":81954153,"identity":"46756fdd-1779-41ca-969c-1f8540593ffe","added_by":"auto","created_at":"2025-05-05 09:41:39","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":237499,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eRibbon Diagrams of Neuraminidase Showing Mutation Locations Relative to Inhibitor Binding Sites (Peramivir, Oseltamivir, Zanamivir, Laninamivir). (A) N44S in PQ836530. (B) I34V, L40I, I106V, and K111R in PQ836531, Highlighted in Red. (C) A20T, Add36H, N44S, V81I, and Other Mutations Across Isolates.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6495442/v1/d1211bfb765aa5f93a6a6a24.png"},{"id":90987164,"identity":"e59377ab-938c-4b6c-a39e-b74e850664bd","added_by":"auto","created_at":"2025-09-10 10:17:17","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1446128,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6495442/v1/60fbf122-6a16-4a98-a339-5a56c1b4adbc.pdf"},{"id":81954156,"identity":"69adb850-aedd-474c-b619-17bef89ef638","added_by":"auto","created_at":"2025-05-05 09:41:39","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1508958,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryFigures.docx","url":"https://assets-eu.researchsquare.com/files/rs-6495442/v1/ee375e2f561e8500b4cd3a86.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Genomic Variability in the Neuraminidase Gene and Antiviral Resistance of Influenza A(H1N1)pdm09 Virus Isolated from Acute Respiratory Infection Patients in Shiraz, Iran (2023–2024)","fulltext":[{"header":"Introduction","content":"\u003cp\u003eInfluenza viruses, members of the Orthomyxoviridae family, are significant global health threats, causing seasonal epidemics and occasional pandemics(\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e). Annually, influenza affects 5 to 15% of the population, leading to 3 to 5\u0026nbsp;million severe cases and up to 650,000 respiratory-related deaths worldwide(\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e). Influenza A viruses, particularly the A(H1N1)pdm09 strain that emerged in 2009, are of major concern due to their high morbidity, mortality, and economic impact(\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e). Shiraz, a major travel hub in Iran, experiences high influenza transmission, yet molecular data are scarce(\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThese single-stranded, negative-sense RNA viruses possess a segmented genome of eight distinct segments encoding essential proteins, including the surface glycoproteins hemagglutinin (HA) and neuraminidase (NA)(\u003cspan additionalcitationids=\"CR6 CR7\" citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e). The NA protein, encoded by the sixth genomic segment, facilitates viral release from infected cells by cleaving sialic acid residues, enabling spread within the respiratory tract(\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e). NA also contributes to viral tropism through its specificity for α-2,6- and α-2,3-linked sialic acids on host cells(\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e). However, the NA gene is highly mutable, undergoing antigenic drift (point mutations) and antigenic shift (genetic reassortment), which drive the emergence of new strains capable of evading immunity and developing antiviral resistance(\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e). Neuraminidase inhibitors (NAIs), such as oseltamivir and zanamivir, are critical for influenza treatment and prophylaxis by blocking NA activity, but resistance\u0026mdash;often linked to mutations like H275Y\u0026mdash;poses a growing challenge(\u003cspan additionalcitationids=\"CR15\" citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eInfluenza A(H1N1)pdm09, responsible for the 2009 pandemic, continues to circulate and evolve, necessitating ongoing molecular surveillance(\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e). Mutations in the NA gene can alter pathogenicity, immune evasion, and drug susceptibility, making their characterization essential for effective influenza management(\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e). This study investigates the genomic changes in the NA gene of A(H1N1)pdm09 isolates from patients with acute respiratory infections in Shiraz, Iran, during 2024\u0026ndash;2025, focusing on antiviral resistance and phylogenetic relationships to inform global surveillance efforts.\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003e\u003cstrong\u003eSample Collection\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBetween November 2023 and December 2024, 300 respiratory samples (nasopharyngeal swabs and sputum) were collected from patients at Shiraz teaching hospitals presenting with acute respiratory infection symptoms, including fever, cough, myalgia, and sore throat. Patients with confirmed co-infections or other respiratory viral infections, identified via multiplex PCR for common respiratory pathogens (e.g., RSV, SARS-CoV-2), were excluded. Samples were immediately placed in viral transport media and stored at -80\u0026deg;C until analysis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical Approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was approved by the Ethics Committee of Shiraz University of Medical Sciences, Shiraz, Iran (approval number: IR.SUMS.MED.REC.1403.557). Informed consent was obtained from all patients or their legal guardians prior to sample collection. All procedures adhered to the Declaration of Helsinki, ensuring participant confidentiality and anonymity throughout the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated and/or analyzed during the current study are available in the \u003cstrong\u003eGenBank\u003c/strong\u003e repository, under the accession numbers \u003cstrong\u003ePQ836530 to PQ836543\u003c/strong\u003e. These sequences can be accessed through the following link: https://www.ncbi.nlm.nih.gov/nuccore/PQ836530-PQ836543.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eRNA Extraction and Real-Time RT-PCR for Detection of Influenza A(H1N1)pdm09\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eViral RNA was extracted from the 300 samples using the High Pure Viral RNA Kit (Roche, Germany) following the manufacturer\u0026rsquo;s protocol, with extracted RNA stored at -80\u0026deg;C to prevent degradation. Real-time RT-PCR was performed using the SuperScript III Platinum One-Step Quantitative RT-PCR Kit (Invitrogen, USA) on a Rotor-Gene 6000 system (Corbett). The reaction mixture (20 \u0026mu;L) consisted of 4 \u0026mu;L RNA, 10 \u0026mu;L 2X master mix, 0.4 \u0026mu;L each of forward and reverse primers (10 \u0026mu;M), 0.4 \u0026mu;L probe (5 \u0026mu;M), and 4.8 \u0026mu;L RNase/DNase-free water. Primers and probes were designed according to the CDC 2024 protocol for Influenza A(H1N1)pdm09 detection (sequences available at [CDC reference or state \u0026lsquo;proprietary\u0026rsquo; if not public]). Thermal cycling conditions were: reverse transcription at 50\u0026deg;C for 30 minutes, enzyme inactivation at 95\u0026deg;C for 2 minutes, followed by 45 cycles of 95\u0026deg;C for 15 seconds, 55\u0026deg;C for 30 seconds, and 72\u0026deg;C for 30 seconds. Of the 300 samples, 14 tested positive for Influenza A(H1N1)pdm09.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAmplification and Sequencing of the Neuraminidase Gene\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eRNA from the 14 positive samples was reverse-transcribed to cDNA using the SuperScript III First-Strand Synthesis System (Invitrogen, USA). The full-length NA gene was amplified via conventional PCR using primers specific for the A(H1N1)pdm09 NA gene (forward: 5\u0026rsquo;-AGCAAAAGCAGGAGT-3\u0026rsquo;, reverse: 5\u0026rsquo;-AGTAGAAACAAGGAGT-3\u0026rsquo;, designed to amplify the 1410 bp NA coding region). PCR reactions (50 \u0026mu;L) contained 5 \u0026mu;L cDNA, 25 \u0026mu;L 2X PCR master mix (TaKaRa, Japan), 1 \u0026mu;L each primer (10 \u0026mu;M), and 18 \u0026mu;L nuclease-free water. Cycling conditions were: 95\u0026deg;C for 3 minutes, followed by 35 cycles of 95\u0026deg;C for 30 seconds, 55\u0026deg;C for 30 seconds, and 72\u0026deg;C for 90 seconds, with a final extension at 72\u0026deg;C for 5 minutes. Amplicons were verified on a 2% agarose gel, purified using the QIAquick PCR Purification Kit (Qiagen, Germany), and sequenced by TopazGene (Iran) using Sanger sequencing.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePhylogenetic and Mutation Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNA sequences were aligned with the reference strain A/California/07/2009 (GenBank: NC_026434.1) using ClustalW in MEGA 11 software. Phylogenetic trees were constructed via the maximum-likelihood method with the Tamura-Nei model, supported by 1000 bootstrap replicates. The 14 NA sequences were deposited in GenBank under accession numbers PQ836530 to PQ836543. Amino acid substitutions were identified by comparing the sequences to A/California/07/2009, focusing on positions associated with antiviral resistance, particularly H275Y for oseltamivir resistance.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStructural Modeling and Antiviral Resistance Assessment\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe NA protein structure was modeled using I-TASSER (C-score: -2.05, TM-score: 0.47\u0026plusmn;0.15). Physicochemical properties were calculated with ProtParam (ExPASy). Molecular docking was performed using AutoDock Vina (v1.1.2) to assess the impact of mutations on inhibitor binding. Ligands (oseltamivir, zanamivir, peramivir, laninamivir) were retrieved from PubChem, converted to PDBQT format using Open Babel (v2.3.1), and docked against the NA model with a grid box centered on the active site (size: 30\u0026times;30\u0026times;30 \u0026Aring;, exhaustiveness: 8). Binding poses were visualized in PyMOL (v2.5).\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eDetection of Influenza A(H1N1)pdm09 in Respiratory Samples\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFrom 300 throat and nasal swab samples collected between November 2023 and December 2024, 14 tested positive for Influenza A(H1N1)pdm09 via Real-Time PCR, confirmed by RT-PCR. The patients, aged 35 to 85 years, included 9 males and 5 females (Table 1), all of whom required hospitalization due to severe respiratory symptoms, including fever, cough, myalgia, and sore throat.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMolecular Characterization of Neuraminidase Gene\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe NA sequences of the 14 A(H1N1)pdm09 isolates were submitted to GenBank under accession numbers PQ836530 to PQ836543 (Table 1). Phylogenetic analysis revealed that these isolates share high nucleotide similarity with the reference strain A/California/07/2009, clustering closely with 2008\u0026ndash;2009 global strains from Luxembourg and Yamagata (Fig. 1).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAmino Acid Variations in Neuraminidase\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSequence analysis identified 26 amino acid substitutions in the NA gene compared to A/California/07/2009, with N44S, I106V, and N200S being the most frequent, observed in 3, 4, and 3 isolates, respectively (Table 2). Other mutations included A20T, I34V, Add36H, L40I, V81I, S90P, R107A, K111R, K143E, I241V, F256L, V264I, N270K, F306I, I314M, I321V, K369N, N386K, N386S, I389T, V394I, K432E, and D451G (Table 3). No isolates exhibited the H275Y mutation associated with oseltamivir resistance, indicating retained sensitivity to neuraminidase inhibitors oseltamivir and zanamivir.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStructural Analysis of Neuraminidase Mutations\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe NA protein was modeled using I-TASSER (C-score: -2.05, TM-score: 0.47\u0026plusmn;0.15). Its physicochemical properties, calculated via ProtParam, showed a theoretical pI of 6.11, an instability index of 32.91, and an aliphatic index of 74.16, indicating moderate stability (Table 4). Molecular docking with inhibitors (peramivir, oseltamivir, zanamivir, and laninamivir) revealed that key mutations, including N44S, I34V, I106V, and others, are located distantly from the inhibitor-binding sites (Fig. 2), suggesting minimal impact on drug efficacy. Superimposition of the I-TASSER model with the 2hu4 crystal structure confirmed structural conservation in the catalytic head domain (Supplementary Fig. 1)\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eInfluenza A(H1N1)pdm09 remains a significant public health concern due to its potential for rapid evolution and antiviral resistance, necessitating continuous molecular surveillance(19). This study identified 26 amino acid substitutions in the NA gene of 14 A(H1N1)pdm09 isolates from Shiraz, Iran, collected between November 2023 and December 2024. The most frequent mutations\u0026mdash;N44S, I106V, and N200S\u0026mdash;were detected in 3, 4, and 3 isolates, respectively, but did not confer resistance to oseltamivir or zanamivir, as no H275Y mutations were observed. This aligns with prior studies in Shiraz from 2012\u0026ndash;2013, which reported no oseltamivir resistance, though a 2016 study identified two resistant isolates with the H275Y mutation(20). Globally, resistance patterns vary; for instance, 34.3% of isolates in Argentina (2008) and 1.4% in Mexico (2009\u0026ndash;2010) exhibited oseltamivir resistance due to H275Y, highlighting the importance of regional surveillance(20, 21).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003ePhylogenetic analysis revealed that our isolates are closely related to the reference strain A/California/07/2009, clustering with 2008\u0026ndash;2009 global strains from Luxembourg and Yamagata. This conserved lineage suggests limited antigenic drift in the NA gene over the past 15 years, consistent with findings from Australia (2001\u0026ndash;2005) where oseltamivir resistance emerged sporadically without widespread drift(22). However, the presence of mutations like N44S and I106V, which are distal from the NA active site as shown by molecular docking, indicates ongoing genetic variability that could influence future pathogenicity or immune evasion(23). Similar distal mutations were reported in Shiraz in 2016 (e.g., V241I, N369K), suggesting a recurring pattern of variation in this region(20).\u003c/p\u003e\n\u003cp\u003eThe absence of oseltamivir resistance in our isolates supports the continued efficacy of NA inhibitors for managing A(H1N1)pdm09 infections in Shiraz. However, the genetic diversity observed underscores the need for vigilance, as mutations near functional sites could eventually impact inhibitor binding, as seen with the S247N mutation reported in other studies(22). Computational modeling, despite limitations such as incomplete loop representation, provides a rapid tool for assessing mutation impacts, which is critical for timely antiviral development(24). Future studies should integrate experimental validation (e.g., enzymatic assays) to confirm the functional effects of these mutations and extend surveillance to other viral proteins like hemagglutinin to fully understand immune escape potential.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study demonstrates significant genetic variability in the NA gene of Influenza A(H1N1)pdm09 isolates from Shiraz, Iran, with 26 mutations identified, though none conferred resistance to oseltamivir or zanamivir. The phylogenetic similarity to A/California/07/2009 highlights a conserved lineage, while the absence of the H275Y mutation supports the ongoing effectiveness of NA inhibitors in this region. These findings emphasize the importance of continuous molecular surveillance to detect emerging mutations that could affect influenza management globally, informing antiviral strategies and public health responses.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\n\u003cp\u003eAmirReza Radfar primarily contributed to the manuscript writing, including the design of the study, data analysis, and interpretation. Mojtaba Mortazavi provided expertise in biotechnology, specifically assisting with the bioinformatics analysis, including structural modeling and molecular docking studies. Mohammad Javad Shahbazi Nezhad, Neda Pirbonyeh, Jamal Sarvari, and Afagh Moattari were responsible for the virology aspects of the study, including sample collection, virological testing, and the molecular characterization of Influenza A(H1N1)pdm09 strains. Afagh Moattari also contributed significantly to the manuscript writing, particularly in the interpretation of the results and the revision of the manuscript. All authors contributed to the drafting and critical revision of the manuscript and approved the final version for submission.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eKumari R, Sharma SD, Kumar A, Ende Z, Mishina M, Wang Y, et al. Antiviral approaches against influenza virus. Clinical microbiology reviews. 2023;36(1):e00040-22.\u003c/li\u003e\n\u003cli\u003eEDITION T. The global impact of respiratory.\u003c/li\u003e\n\u003cli\u003eMahmud S, Tamanna RK. Influenza Virus Types, Subtypes and Genomic Lineage with Its Prevention and Control: A Narrative Review. Journal of Monno Medical College. 2024;10(1):43-51.\u003c/li\u003e\n\u003cli\u003eFaramarzi H, Mousavi-Roknabadi RS, Hemmati A, Faramarzi A, Bakhtiari H. Epidemiology of Influenza in Fars Province, Southern Iran; a Population-Based Study (2015\u0026ndash;2019). Archives of Iranian Medicine. 2021;24(3):199-208.\u003c/li\u003e\n\u003cli\u003eNoda T. Selective genome packaging mechanisms of influenza A viruses. Cold Spring Harbor Perspectives in Medicine. 2021;11(7):a038497.\u003c/li\u003e\n\u003cli\u003eFlerlage T, Boyd DF, Meliopoulos V, Thomas PG, Schultz-Cherry S. Influenza virus and SARS-CoV-2: pathogenesis and host responses in the respiratory tract. Nature Reviews Microbiology. 2021;19(7):425-41.\u003c/li\u003e\n\u003cli\u003eRussell CJ. Hemagglutinin stability and its impact on influenza A virus infectivity, pathogenicity, and transmissibility in avians, mice, swine, seals, ferrets, and humans. Viruses. 2021;13(5):746.\u003c/li\u003e\n\u003cli\u003eMoattari A, Emami A, Moghadami M, Honarvar B. Influenza viral infections among the Iranian Hajj pilgrims returning to Shiraz, Fars province, Iran. Influenza and other respiratory viruses. 2012;6(6):e77-e9.\u003c/li\u003e\n\u003cli\u003eLi Y, Liu D, Wang Y, Su W, Liu G, Dong W. The importance of glycans of viral and host proteins in enveloped virus infection. Frontiers in Immunology. 2021;12:638573.\u003c/li\u003e\n\u003cli\u003eChauhan RP, Gordon ML. An overview of influenza A virus genes, protein functions, and replication cycle highlighting important updates. Virus genes. 2022;58(4):255-69.\u003c/li\u003e\n\u003cli\u003eBurzyńska P, Sobala ŁF, Mikołajczyk K, Jodłowska M, Jaśkiewicz E. Sialic acids as receptors for pathogens. Biomolecules. 2021;11(6):831.\u003c/li\u003e\n\u003cli\u003ePerofsky AC, Huddleston J, Hansen CL, Barnes JR, Rowe T, Xu X, et al. Antigenic drift and subtype interference shape A (H3N2) epidemic dynamics in the United States. Elife. 2024;13:RP91849.\u003c/li\u003e\n\u003cli\u003eDapat C, Saito R, Kyaw Y, Myint YY, Oo HN, Oo KY, et al. Delayed emergence of oseltamivir-resistant seasonal influenza A (H1N1) and pandemic influenza A(H1N1)pdm09 viruses in Myanmar. Influenza Other Respir Viruses. 2013;7(5):766-71.\u003c/li\u003e\n\u003cli\u003eYang JR, Huang YP, Chang FY, Hsu LC, Huang HY, Pan YT, et al. Characterization of oseltamivir‐resistant influenza A (H1N1) pdm09 viruses in Taiwan in 2009\u0026ndash;2011. Journal of Medical Virology. 2013;85(3):379-87.\u003c/li\u003e\n\u003cli\u003eRussell RJ, Haire LF, Stevens DJ, Collins PJ, Lin YP, Blackburn GM, et al. The structure of H5N1 avian influenza neuraminidase suggests new opportunities for drug design. Nature. 2006;443(7107):45-9.\u003c/li\u003e\n\u003cli\u003eGoka E, Vallely P, Mutton K, Klapper P. Mutations associated with severity of the pandemic influenza A (H1N1) pdm09 in humans: a systematic review and meta-analysis of epidemiological evidence. Archives of Virology. 2014;159:3167-83.\u003c/li\u003e\n\u003cli\u003eMedina RA, Garc\u0026iacute;a-Sastre A. Influenza A viruses: new research developments. Nature Reviews Microbiology. 2011;9(8):590-603.\u003c/li\u003e\n\u003cli\u003eMcAuley JL, Gilbertson BP, Trifkovic S, Brown LE, McKimm-Breschkin JL. Influenza virus neuraminidase structure and functions. Frontiers in microbiology. 2019;10:39.\u003c/li\u003e\n\u003cli\u003eWang X, Xiao F, Wang Y, Deng X, Chen Z, Dong X, et al. Changed epidemiology of narcolepsy before, during, and after the 2009 H1N1 pandemic: a nationwide narcolepsy surveillance network study in mainland China, 1990\u0026ndash;2017. Sleep. 2023;46(3):zsac325.\u003c/li\u003e\n\u003cli\u003eDia N, Niang MN, Diadhiou SA, Goudiaby DG, Faye A, Kiori D, et al. Spread of Influenza A (H1N1) oseltamivir-resistant viruses in Africa in 2008 confirmed by multiple introductions in Senegal. BMC Infectious Diseases. 2013;13:1-7.\u003c/li\u003e\n\u003cli\u003ePontoriero A, Baumeister E, Campos AM, Savy VL. Vigilancia virologica y resistencia a los antiviricos del virus de la gripe humana en la Argentina, 2005-2008. Revista Panamericana de Salud Publica. 2011;30:634+.\u003c/li\u003e\n\u003cli\u003eTamura D, DeBiasi RL, Okomo-Adhiambo M, Mishin VP, Campbell AP, Loechelt B, et al. Emergence of Multidrug-Resistant Influenza A(H1N1)pdm09 Virus Variants in an Immunocompromised Child Treated With Oseltamivir and Zanamivir. The Journal of Infectious Diseases. 2015;212(8):1209-13.\u003c/li\u003e\n\u003cli\u003eNguyen TH, Anh Pham NQ, Thai QM, Vu VV, Ngo ST, Horng J-T. Discovering Influenza Virus Neuraminidase Inhibitors via Computational and Experimental Studies. ACS Omega. 2024;9(49):48505-11.\u003c/li\u003e\n\u003cli\u003eVavricka CJ, Liu Y, Kiyota H, Sriwilaijaroen N, Qi J, Tanaka K, et al. Influenza neuraminidase operates via a nucleophilic mechanism and can be targeted by covalent inhibitors. Nat Commun. 2013;4:1491.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":" \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cdiv class=\"SimplePara\"\u003eDemographic and Genetic Details of Influenza A(H1N1)pdm09 Strains\u003c/div\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eSex\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eGene Bank Accession\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cdiv class=\"SimplePara\"\u003eOrigin\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cdiv class=\"SimplePara\"\u003eInfluenza A(H1N1) pdm09 Strains Name\u003c/div\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eF\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eM\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eM\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eM\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eF\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eM\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eF\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eM\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eM\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eM\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eF\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eM\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eM\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eM\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836530\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836531\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836532\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836533\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836534\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836535\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836536\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836537\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836538\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836539\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836540\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836541\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836542\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836543\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cdiv class=\"SimplePara\"\u003eShiraz\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eShiraz\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eShiraz\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eShiraz\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eShiraz\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eShiraz\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eShiraz\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eShiraz\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eShiraz\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eShiraz\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eShiraz\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eShiraz\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eShiraz\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eShiraz\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cdiv class=\"SimplePara\"\u003eInfluenza A virus (A/Iran/01/2024(H1N1))\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eInfluenza A virus (A/Iran/02/2024(H1N1))\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eInfluenza A virus (A/Iran/03/2024(H1N1))\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eInfluenza A virus (A/Iran/04/2024(H1N1))\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eInfluenza A virus (A/Iran/05/2024(H1N1))\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eInfluenza A virus (A/Iran/06/2024(H1N1))\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eInfluenza A virus (A/Iran/07/2024(H1N1))\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eInfluenza A virus (A/Iran/08/2024(H1N1))\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eInfluenza A virus (A/Iran/09/2024(H1N1))\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eInfluenza A virus (A/Iran/10/2024(H1N1))\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eInfluenza A virus (A/Iran/11/2024(H1N1))\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eInfluenza A virus (A/Iran/12/2024(H1N1))\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eInfluenza A virus (A/Iran/13/2024(H1N1))\u003c/div\u003e \u003cdiv class=\"SimplePara\"\u003eInfluenza A virus (A/Iran/14/2024(H1N1))\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003cbr/\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cdiv class=\"SimplePara\"\u003eAmino Acid Variation Across Influenza A(H1N1)pdm09 Isolates\u003c/div\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"27\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c14\" colnum=\"14\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c15\" colnum=\"15\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c16\" colnum=\"16\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c17\" colnum=\"17\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c18\" colnum=\"18\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c19\" colnum=\"19\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c20\" colnum=\"20\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c21\" colnum=\"21\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c22\" colnum=\"22\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c23\" colnum=\"23\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c24\" colnum=\"24\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c25\" colnum=\"25\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c26\" colnum=\"26\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c27\" colnum=\"27\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cdiv class=\"SimplePara\"\u003eIsolates\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"26\" nameend=\"c27\" namest=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eNumber of amino acid\u003c/div\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003e20\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cdiv class=\"SimplePara\"\u003e34\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cdiv class=\"SimplePara\"\u003e36\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cdiv class=\"SimplePara\"\u003e40\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cdiv class=\"SimplePara\"\u003e44\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cdiv class=\"SimplePara\"\u003e81\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cdiv class=\"SimplePara\"\u003e90\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cdiv class=\"SimplePara\"\u003e106\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cdiv class=\"SimplePara\"\u003e107\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cdiv class=\"SimplePara\"\u003e111\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c12\"\u003e \u003cdiv class=\"SimplePara\"\u003e143\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c13\"\u003e \u003cdiv class=\"SimplePara\"\u003e200\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c14\"\u003e \u003cdiv class=\"SimplePara\"\u003e241\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c15\"\u003e \u003cdiv class=\"SimplePara\"\u003e256\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c16\"\u003e \u003cdiv class=\"SimplePara\"\u003e264\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c17\"\u003e \u003cdiv class=\"SimplePara\"\u003e270\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c18\"\u003e \u003cdiv class=\"SimplePara\"\u003e306\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c19\"\u003e \u003cdiv class=\"SimplePara\"\u003e314\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c20\"\u003e \u003cdiv class=\"SimplePara\"\u003e321\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c21\"\u003e \u003cdiv class=\"SimplePara\"\u003e369\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c22\"\u003e \u003cdiv class=\"SimplePara\"\u003e386\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c23\"\u003e \u003cdiv class=\"SimplePara\"\u003e386\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c24\"\u003e \u003cdiv class=\"SimplePara\"\u003e389\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c25\"\u003e \u003cdiv class=\"SimplePara\"\u003e394\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c26\"\u003e \u003cdiv class=\"SimplePara\"\u003e432\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c27\"\u003e \u003cdiv class=\"SimplePara\"\u003e451\u003c/div\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eA/California/07/2009\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eA/shiraz/01\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cdiv class=\"SimplePara\"\u003eS\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eA/shiraz/02\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cdiv class=\"SimplePara\"\u003eV\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cdiv class=\"SimplePara\"\u003eI\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cdiv class=\"SimplePara\"\u003eS\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cdiv class=\"SimplePara\"\u003eV\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cdiv class=\"SimplePara\"\u003eR\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cdiv class=\"SimplePara\"\u003eS\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cdiv class=\"SimplePara\"\u003eI\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e \u003cdiv class=\"SimplePara\"\u003eK\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e \u003cdiv class=\"SimplePara\"\u003eM\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e \u003cdiv class=\"SimplePara\"\u003eV\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e \u003cdiv class=\"SimplePara\"\u003eK\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e \u003cdiv class=\"SimplePara\"\u003eE\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eA/shiraz/03\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cdiv class=\"SimplePara\"\u003eV\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e \u003cdiv class=\"SimplePara\"\u003eN\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cdiv class=\"SimplePara\"\u003eI\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eA/shiraz/04\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eT\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cdiv class=\"SimplePara\"\u003eH\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cdiv class=\"SimplePara\"\u003eP\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e \u003cdiv class=\"SimplePara\"\u003eI\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eA/shiraz/05\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cdiv class=\"SimplePara\"\u003eI\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eA/shiraz/06\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cdiv class=\"SimplePara\"\u003eH\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e \u003cdiv class=\"SimplePara\"\u003eS\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e \u003cdiv class=\"SimplePara\"\u003eG\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eA/shiraz/07\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cdiv class=\"SimplePara\"\u003eH\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cdiv class=\"SimplePara\"\u003eS\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cdiv class=\"SimplePara\"\u003eV\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cdiv class=\"SimplePara\"\u003eS\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e \u003cdiv class=\"SimplePara\"\u003eI\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eA/shiraz/08\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eA/shiraz/09\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cdiv class=\"SimplePara\"\u003eH\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eA/shiraz/10\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cdiv class=\"SimplePara\"\u003eA\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eA/shiraz/11\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cdiv class=\"SimplePara\"\u003eH\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cdiv class=\"SimplePara\"\u003eS\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cdiv class=\"SimplePara\"\u003eV\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cdiv class=\"SimplePara\"\u003eE\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cdiv class=\"SimplePara\"\u003eS\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eA/shiraz/12\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eA/shiraz/13\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cdiv class=\"SimplePara\"\u003eS\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eA/shiraz/14\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cdiv class=\"SimplePara\"\u003eH\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cdiv class=\"SimplePara\"\u003eS\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cdiv class=\"SimplePara\"\u003eV\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cdiv class=\"SimplePara\"\u003eS\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cdiv class=\"SimplePara\"\u003eL\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c17\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c18\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c19\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c20\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c21\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c22\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c23\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c24\"\u003e \u003cdiv class=\"SimplePara\"\u003eT\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c25\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c26\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c27\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003cbr/\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cdiv class=\"SimplePara\"\u003eMutations in Influenza A(H1N1)pdm09 Isolates by Gene Bank Accession\u003c/div\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"2\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eGene Bank Accession\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eMutation\u003c/div\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836530\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eN44S\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836531\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eI34V, L401, I106V, K111R\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836532\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eI241V, K369N\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836533\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eA20T, S90P, F306I\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836534\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eV81I\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836535\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eN386S\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836536\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eV394I\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836538\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eAdd 36H\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836539\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eR107A\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836540\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eK143E, N200S\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836542\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eN44S\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003ePQ836543\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eF256L, I389T\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003cbr/\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cdiv class=\"SimplePara\"\u003eBiochemical and Structural Parameters of Neuraminidase in Influenza A(H1N1)pdm09 Isolates\u003c/div\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"2\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eParameter\u003c/div\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003eValue\u003c/div\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eTheoretical pI\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003e6.11\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eMolecular weight\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003e51613.01\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eSequence length\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003e469\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eExtinction coefficients (M-1 cm-1 at 260 nm)*\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003e109985\u0026thinsp;\u0026minus;\u0026thinsp;108860\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eAsp\u0026thinsp;+\u0026thinsp;Glu\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003e40\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eArg\u0026thinsp;+\u0026thinsp;Lys\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003e37\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eInstability index\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003e32.91\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eGrand average of hydropathicity\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003e-0.259\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cdiv class=\"SimplePara\"\u003eAliphatic index\u003c/div\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cdiv class=\"SimplePara\"\u003e74.16\u003c/div\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e "}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Neuraminidase, Influenza A(H1N1)pdm09, oseltamivir resistance, mutation, phylogenetic analysis, molecular surveillance","lastPublishedDoi":"10.21203/rs.3.rs-6495442/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6495442/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eThe 2009 Influenza A(H1N1)pdm09 pandemic caused significant global morbidity and mortality. Mutations in the neuraminidase (NA) gene can influence viral pathogenicity, immune evasion, and resistance to antiviral drugs, necessitating continuous molecular surveillance to guide antiviral development.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003eFrom November 2023 to December 2024, 300 respiratory samples were collected from patients with acute respiratory infections in Shiraz, Iran. Real-Time PCR identified Influenza A(H1N1)pdm09, followed by conventional PCR amplification and sequencing of the NA gene. Mutations and resistance to neuraminidase inhibitors (oseltamivir, zanamivir) were assessed using MEGA 11, I-TASSER modeling, and AutoDock Vina docking. Phylogenetic analysis was performed to evaluate genetic relationships.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eOf the 300 samples, 14 were positive for Influenza A(H1N1)pdm09. Sequence analysis identified 26 NA gene mutations, with N44S, I106V, and N200S detected in 3, 4, and 3 isolates, respectively. No isolates showed the H275Y mutation, indicating sensitivity to oseltamivir and zanamivir. Phylogenetically, the isolates were highly similar to A/California/07/2009, clustering with 2008\u0026ndash;2009 global strains from Luxembourg and Yamagata.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eDespite significant genetic variability in the NA gene, current antiviral therapies remain effective against these isolates. This study underscores the importance of ongoing surveillance to detect emerging mutations that could impact influenza management globally.\u003c/p\u003e","manuscriptTitle":"Genomic Variability in the Neuraminidase Gene and Antiviral Resistance of Influenza A(H1N1)pdm09 Virus Isolated from Acute Respiratory Infection Patients in Shiraz, Iran (2023–2024)","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-05 09:41:35","doi":"10.21203/rs.3.rs-6495442/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"0fc89964-5ddf-4e92-a34b-80540b489804","owner":[],"postedDate":"May 5th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":47715407,"name":"Biological sciences/Biotechnology"},{"id":47715408,"name":"Biological sciences/Microbiology"}],"tags":[],"updatedAt":"2025-09-10T10:09:04+00:00","versionOfRecord":[],"versionCreatedAt":"2025-05-05 09:41:35","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6495442","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6495442","identity":"rs-6495442","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}
Text is read by the "Ask this paper" AI Q&A widget below. Extraction quality varies by source — PMC NXML preserves structure cleanly, OA-HTML may include some navigation residue, and OA-PDF can have broken hyphenation. The publisher copy (via DOI) is the canonical version.
My notes (saved in your browser only)
Ask this paper
Answers must be backed by verbatim quotes from this paper's full text. Hallucinated quotes are dropped automatically; if no verbatim passage answers the question, we say so. How this works
Citation neighborhood (no data yet)
We don't have any in-corpus citations linked to this paper yet. This is a recent paper (2025) — citers typically take a year or two to land, and the OpenAlex reference graph may still be filling in.
Source provenance
- europepmc
- last seen: 2026-05-20T01:45:00.602351+00:00
- unpaywall
- last seen: 2026-05-22T02:00:06.705733+00:00
License: CC-BY-4.0