agr Genotyping and Biofilm Formation in Staphylococci Isolated From Clinical Specimens

agr Genotyping and Biofilm Formation in Staphylococci Isolated From Clinical Specimens | 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 agr Genotyping and Biofilm Formation in Staphylococci Isolated From Clinical Specimens Nilofor Ansari, Sarita Manandhar, Anil Kumar Sah This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8630905/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 Abstract Staphylococci, a type of Gram-positive cocci, are frequently associated with invasive infections, posing difficulties for effective treatment due to various virulence factors. The accessory gene regulator (agr) gene and biofilm production are known contributors to these treatment challenges. The purpose of this study was to examine the presence of biofilm- forming strains, the agr gene, and patterns of multidrug resistance in staphylococci. A total of 363 staphylococci were isolated from various clinical specimens at Annapurna Neurological and Allied Science Hospital, Maitighar. Kathmandu from August 2023 to February 2024. Disk diffusion test was performed for the methicillin resistant S. aureus (MRSA) and coagulase negative staphylococci (CONS). The antibiotic susceptibility test was performed by Kirby Bauer disc diffusion method following CLSI guidelines. To identify clindamycin and erythromycin resistance a D-test was performed by Kirby Bauer disc diffusion method. Detection of biofilm formation was performed by microtitre plate assay followed by Polymerase Chain Reaction for agr genotyping. Among total isolates 82.9% were S. aureus and 17% were CONS. The disc diffusion method detected 17% of isolates were MRCONS and 36.58% of isolates were MRSA isolates. Both MRSA and MRCONS showed higher resistance towards erythromycin and ampicillin. MRCONS showed multidrug resistance. Only one Methicillin Susceptible Staphylococcus aureus (MSSA) isolate (5.5%) showed Susceptible pattern, 3 MSSA isolates (16.67%) isolates showed Inducible Macrolide- Lincosamide- Streptogramin B (IMLSB) pattern and 2 (11.11%) isolates showed Macrolide- Lincosamide- Streptogramin B (MSB) pattern. Similarly, among MRSA 3 (18.75%) isolates showed Constitutive Macrolide- Lincosamide- Sterptog-ramin B (CMLSB) pattern, 1 (6.25%) isolate showed susceptible pattern and 2 (12.5%) isolates showed IMLSB pattern. A total of 14 (34.15%) isolates were biofilm producers and 27 (65.85%) were biofilm non-producers. Among biofilm producers higher resistance was found against erythromycin (57.1%). Out of total isolates one isolate was agr I (9.1%) positive and 10 (24.4%) isolates were agr III positive. The current investigation revealed that S. aureus and CONS were highly prevalent in the hospital environment. Staphylococcal isolates have demonstrated resistance to (beta) β- lactams and macrolides, which is consistent with the trend of the multiple drugs resistance emerging. Among biofilm formers S. aureus has shown higher resistance towards antibiotics than CONS. Among the isolates agr I and agr III gene has been discovered. The percentage of MRSA from our region that carry this gene seems to be rather high based on the prevalence of the agr gene in MRSA. Health sciences/Diseases Biological sciences/Microbiology MRCONS MRSA agr biofilm producer biofilm non-producer Full Text Additional Declarations No competing interests reported. Supplementary Files Supplementaryfile.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-8630905","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":577542265,"identity":"7c9ddb2d-31bd-4924-849e-0ba81fce6637","order_by":0,"name":"Nilofor 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\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"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":"MRCONS, MRSA, agr, biofilm producer, biofilm non-producer ","lastPublishedDoi":"10.21203/rs.3.rs-8630905/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8630905/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eAbstract Staphylococci, a type of Gram-positive cocci, are frequently associated with invasive infections, posing difficulties for effective treatment due to various virulence factors. The accessory gene regulator (agr) gene and biofilm production are known contributors to these treatment challenges. The purpose of this study was to examine the presence of biofilm- forming strains, the agr gene, and patterns of multidrug resistance in staphylococci. A total of 363 staphylococci were isolated from various clinical specimens at Annapurna Neurological and Allied Science Hospital, Maitighar. Kathmandu from August 2023 to February 2024.\u0026nbsp; Disk diffusion test was performed for the methicillin resistant S. aureus (MRSA) and coagulase negative staphylococci (CONS). The antibiotic susceptibility test was performed by Kirby Bauer disc diffusion method following CLSI guidelines. To identify clindamycin and erythromycin resistance a D-test was performed by Kirby Bauer disc diffusion method.\u0026nbsp; Detection of biofilm formation was performed by microtitre plate assay followed by Polymerase Chain Reaction for agr genotyping.\u0026nbsp; Among total isolates 82.9% were S. aureus and 17% were CONS. The disc diffusion method detected 17% of isolates were MRCONS and 36.58% of isolates were MRSA isolates. Both MRSA and MRCONS showed higher resistance towards erythromycin and ampicillin.\u0026nbsp; MRCONS showed multidrug resistance. Only one Methicillin Susceptible Staphylococcus aureus (MSSA) isolate (5.5%) showed Susceptible pattern, 3 MSSA isolates (16.67%) isolates showed Inducible Macrolide- Lincosamide- Streptogramin B (IMLSB) pattern and 2 (11.11%) isolates showed Macrolide- Lincosamide- Streptogramin B (MSB) pattern. Similarly, among MRSA 3 (18.75%) isolates showed Constitutive Macrolide- Lincosamide- Sterptog-ramin B (CMLSB) pattern, 1 (6.25%) isolate showed susceptible pattern and 2 (12.5%) isolates showed IMLSB pattern. A total of 14 (34.15%) isolates were biofilm producers and 27 (65.85%) were biofilm non-producers. Among biofilm producers higher resistance was found against erythromycin (57.1%). Out of total isolates one isolate was agr I (9.1%) positive and 10 (24.4%) isolates were agr III positive. The current investigation revealed that S. aureus and CONS were highly prevalent in the hospital environment. Staphylococcal isolates have demonstrated resistance to (beta) β- lactams and macrolides, which is consistent with the trend of the multiple drugs resistance emerging. Among biofilm formers S. aureus has shown higher resistance towards antibiotics than CONS.\u0026nbsp; Among the isolates agr I and agr III gene has been discovered. The percentage of MRSA from our region that carry this gene seems to be rather high based on the prevalence of the agr gene in MRSA.\u003c/p\u003e","manuscriptTitle":"agr Genotyping and Biofilm Formation in Staphylococci Isolated From Clinical Specimens","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-01-22 04:32:29","doi":"10.21203/rs.3.rs-8630905/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":"873a1ab7-af24-44d8-bf48-211e94cd2558","owner":[],"postedDate":"January 22nd, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":61549225,"name":"Health sciences/Diseases"},{"id":61549226,"name":"Biological sciences/Microbiology"}],"tags":[],"updatedAt":"2026-03-23T10:27:47+00:00","versionOfRecord":[],"versionCreatedAt":"2026-01-22 04:32:29","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8630905","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8630905","identity":"rs-8630905","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}