Genomic Insights and Recent Epidemiology of a Multidrug-Resistant Mycoplasma hominis Isolate in China: Implications for Clinical Management

Genomic Insights and Recent Epidemiology of a Multidrug-Resistant Mycoplasma hominis Isolate in China: Implications for Clinical Management | 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 Short Report Genomic Insights and Recent Epidemiology of a Multidrug-Resistant Mycoplasma hominis Isolate in China: Implications for Clinical Management Hangfei Chen, Zhenping Wu, Qing Yang, Wenqiao Yu, Hua Zhou This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9183112/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 12 You are reading this latest preprint version Abstract Mycoplasma hominis (M. hominis) is an opportunistic pathogen linked to urogenital and neonatal infections; however, limited genetic and epidemiological data are available. Extragenital infections in healthy adults are rare, and effective antibiotics are species-specific, complicating diagnosis and treatment. Hence, this study aimed to elucidate the clinical process, update the epidemiological characteristics, and investigate the genomic features of a multidrug-resistant M. hominis isolate from an immunocompetent pleuropneumonia patient in China. The M. hominis isolate ZY_MH01 was recovered from pleural fluid and was identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and Whole Genome Sequencing (WGS). The completed genome was annotated using the NCBI Prokaryotic Genome Annotation Pipeline (PGAP). Snippy v4.4.5 was utilized to conduct a core genome single nucleotide polymorphism (cgSNP) analysis between ZY_MH01 and 50 M. hominis strains from the NCBI GenBank database. Subsequently, phylogenies were constructed using IQtree v2.0.3 and visualized by iTOL. Antimicrobial resistance genes and susceptibility were identified by CARD RGI v6.0.3 and VITEK 2 Compact System. The antimicrobial resistance mutation in gene parC (S91I) was found in the genome. Phylogenetic analysis revealed that ZY_MH01 and strains from the NCBI database were epidemiologically related, with ZY_MH01's closest relatives being in the same city and sharing the same resistance and virulence genes. M. hominis should be considered a potential cause of pulmonary infection, particularly in patients unresponsive to broad-spectrum antibiotics. This study provides a detailed analysis of M. hominis epidemiology, highlighting regional evolutionary relationships among strains. Mycoplasma hominis pneumonia immunocompetent multidrug-resistance cgSNP Figures Figure 1 Figure 2 Figure 3 Introduction M. hominis belongs to the genus Mycoplasma, which is commonly classified into two categories: genital and respiratory Mycoplasmas. The former category encompasses M. hominis and Ureaplasma urealyticum , as well as Ureaplasma parvum , all of which are frequently detected in the human urogenital tract in both symptomatic patients and healthy individuals. However, apart from a few cases that lead to genital tract infections and neonatal infections 1 , most individuals do not develop any disease 2 . In recent decades, multiple cases of extra-genital infections have been reported, including postoperative site infections such as osteitis and arthritis, mediastinitis, endocarditis, peritonitis, pyelonephritis, brain abscess, pneumonia, and wound infections, mainly in immunosuppressed patients. In addition, similar cases have also been described in immunocompetent patients, particularly in individuals with predisposing factors such as surgery and trauma. Respiratory infection with M. hominis is relatively rare and primarily occurs in the form of disseminated infection in immunocompromised patients or patients with altered cardiorespiratory function 3 ~ 5 . In this study, the M. hominis strain ZY_MH01 was collected from an immunocompetent patient with respiratory M. hominis infection after blunt chest trauma. Additionally, the case's clinical features, imaging characteristics, and therapeutic options were analyzed by reviewing domestic and international literature. A 47-year-old male fell from a height of eight meters and suffered bilateral multiple rib fractures and traumatic hemopneumothorax. The patient was previously healthy and his past medical history was insignificant. Chest Computed Tomography (CT) examination exhibited pelvic fractures, bilateral multiple rib fractures, and slight pleural effusion accompanied by atelectasis. The patient received thoracic cavity closed drainage and was managed conservatively. Fever and dyspnea appeared in six days. A repeat CT scan revealed more pleural effusion and slight pneumothorax. The patient developed respiratory failure and was intubated and placed under mechanical ventilation. The body temperature remained at 39.5℃; routine blood examination showed a white blood cell (WBC) count of 24.21*10 9 /L, a neutrophil count of 22.85*10 9 /L, C-reactive protein (CRP) levels of 146.73mg/L (normal value < 10 mg/L), and procalcitonin levels of 4.75ng/mL. Empirical therapy was initiated for a suspected bacterial infection: cefoperazone sulbactam sodium (2.0g ivgtt q8h) combined with vancomycin (1g ivgtt q12h) was given with expectorant and other supportive treatment. An additional chest drainage was performed and 200 mL of light red pleural effusion was removed. The patient's body temperature and pulse rate temporarily decreased to 37.5℃ and 63/min, respectively. Subsequently, both parameters increased and remained elevated. A repeat chest CT scan showed patchy and increased density shadows, suggesting the possibility of a rapidly progressing lung infection (Fig. 1 A, B). The results of pleural effusion culture revealed pin-point-sized colonies on Columbia blood agar plates, which were identified as M. hominis by MALDI-TOF MS. Antimicrobial treatment was switched to omadacycline (0.1g ivgtt qd), and the antimicrobial susceptibility testing was determined by VITEK 2 Compact System, demonstrating that the M. hominis strain was resistant to clarithromycin and ciprofloxacin, but sensitive to tetracyclic (Table.1). The patient’s general condition, CRP, and oxygenation gradually returned to normal in the following 7 days, and omadacycline was stopped after total treatment duration of 18 days. The repeat chest CT showed that the majority of niduses returned to normal, with only minor pleural effusion (Fig. 1 C, D). The patient fully recovered and was discharged a few days later. Methods and Results The M. hominis strain ZY_MH01 was further confirmed by WGS. Total genomic DNA was extracted using a QIAamp DNA Mini Kit (Qiagen, Valencia, CA, USA) according to the manufacturer’s instructions. The quality of the extracted DNA was examined by a NanoDrop spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). A DNA library was then prepared using a Nextera DNA Library Preparation Kit (Illumina, San Diego, CA, USA), and WGS was performed on the Illumina HiSeq X10 platform (Illumina, San Diego, CA, USA) and long read Oxford Nanopore platform (Oxford, UK). After sequencing, all sequence reads were pre-processed to remove low-quality or artefactual bases. FastQC v.0.11.8 and Filtlong were separately used to assess the quality of the shotgun and long-read raw data. The clean data have been hybrid assembled by Unicycler. The Whole Genome project has been deposited at DDBJ/ENA/GenBank under the accession JBFWID000000000. Furthermore, the completed genome sequence of M. hominis ZY_MH01 was subsequently assigned using Prokka v1.2.0. The sequence comprises 686,552 bp, with the overall G+C content accounting for 27.1% (Figure 2). Gene prediction was performed using Prodigal in single-genome mode to avoid overprediction commonly observed in small complete bacterial genomes. In total, 604 coding sequences, including 1 tmRNA, 4 rRNA and 33 tRNA genes, were identified. Virulence factors and antibiotic resistance were identified by the online web servers Virulence Factors Database (VFDB)-2022 and Comprehensive Antibiotic Resistance Database (CARD)-2024, which identified that the strain ZY_MH01 contains virulence genes tuf (elongation factor) and hlyA (membrane-associated hemolysin) and antimicrobial resistance mutation in gene parC (S91I) (fluoroquinolone resistant) compare to the reference genome M. hominis ATCC 23114 (GenBank accession no. FP236530.1). The phylogenetic relationship between M. hominis ZY_MH01 and 50 M. hominis strains with comparable sequence assembly levels, currently available in the NCBI GenBank database, was analyzed (Figure 3). These strains were recovered from Russia (21 strains), the UK (20 strains), the USA (5 strains), China (3 strains), and France (1 strain), respectively. The data suggested that these strains were epidemiologically related, and the antimicrobial resistance genes also exhibited regional associations. The closest relative of ZY_MH01 is MH-1, which was also recovered from Hangzhou, China, and this strain shares the same antimicrobial resistance mutation parC (S91I) and virulence genes tuf and hlyA . Discussion and Conclusion M. hominis belongs to the genus Mycoplasma of the Mycoplasmataceae family within the Mollicutes class, which is characterized by the absence of a cell wall. M. hominis is a fastidious slow-growing organism that is hard to detect with routine laboratory methods, including gram staining and routine bacterial culture 6,7 . This study reports a M. hominis strain ZY_MH01 collected from a patient with pleuropneumonia. Due to the characteristic lack of a rigid cell wall, ZY_MH01 is intrinsically resistant to antimicrobials targeting the cell wall, including β-lactam antibiotics and vancomycin, leading to empirical therapy failure. Timely positive bacterial culture results promote quick treatment of the infection. However, M. hominis is difficult to diagnose due to the microbe’s sensitivity to environmental factors and high demands for living conditions, thereby requiring delicate culture methods to yield a positive result 8 . Conventional microbiological tests, such as culture, smear microscopy, and PCR, require prior assumptions. Nonetheless, combining conventional tests with next-generation sequencing, including untargeted metagenomic or targeted approaches, could enhance pathogen diagnosis and further identify epidemiological information, virulence, and resistance genes. The phylogenetic analysis revealed that M. hominis demonstrated epidemiological relatedness, a factor previously deemed irrelevant in earlier research 9 , potentially due to the limited number of reported strains. Additionally, the closest phylogenetic relatives were found to frequently possess identical antimicrobial resistance genes and virulence genes. Both ZY_MH01 and its closest relative MH-1 contain the virulence genes tuf and hlyA , which indicate that their pathogenicity is similar to common strains of Mycoplasma spp 10 . The fluoroquinolone resistance gene parC has been identified in ofloxacin and ciprofloxacin-resistant phenotypes, which is in accordance with a recent meta-analysis, revealing an increase of 59.8% and 31.2% in the resistance rate of ciprofloxacin and ofloxacin, respectively 11 . Moreover, in contrast to other Mycoplasma spp., M. hominis is intrinsically resistant to macrolide antibiotics; hence tetracyclines are recommended as the first-line agents for mycoplasma 2,12 . In summary, M. hominis is not typically considered a common etiological agent of pleuropneumonia, and its identification poses challenges due to the necessity for extended anaerobic culturing. Molecular diagnostic techniques are essential and serve as a critical adjunct to traditional culture methods. Clinicians should remain cognizant of the potential involvement of this organism in a range of infections, particularly in immunocompromised individuals or those with a history of surgical procedures, trauma, or invasive catheterization. These findings enhance the understanding of the pathogen's genomic epidemiology and resistance mechanisms, thereby aiding timely diagnosis and appropriate antibiotic treatment to prevent long-term complications. Abbreviations M. hominis Mycoplasma hominis MALDI-TOF MS matrix-assisted laser desorption/ionization time-of-flight mass spectrometry CT Computed tomography CRP C-reactive protein WBC white blood cell MIC minimum inhibiting concentration Declarations Author Contributions Hua Zhou and Wenqiao Yu were responsible for the organization and coordination of the study. Hangfei Chen was responsible for the data analysis and the finalization of the manuscript. Zhenping Wu served as the chief investigator, and Qtng Yang provided diagnostic assistance. All authors contributed to the writing of the final manuscript. Consent and research ethics This study has been reviewed and approved by the Research Ethics Committee of the First Affiliated Hospital of Zhejiang University (ITT20240876). The patient provided informed consent for publication of the clinical details including lung CT images, and written informed consent was obtained. Disclosure The authors declare that they have no conflicts of interest. Funding This work was supported by research grants, including the Key R&D Plan of the Ministry of Science and Technology of China [grant number 2022YFC2504502]; the Research and Development Program of Zhejiang Province [grant number 2023C03068]; the National Natural Science Foundation of China [grant number 82272338], awarded to Hua Zhou. The funders were not involved in the study design, data collection and analysis, the decision to publish, or the preparation of the manuscript. References Capoccia R, Greub G, Baud D. Ureaplasma urealyticum, Mycoplasma hominis and adverse pregnancy outcomes. Current opinion in infectious diseases. 2013;26(3):231-240. Machalek DA, Tao Y, Shilling H, et al. Prevalence of mutations associated with resistance to macrolides and fluoroquinolones in Mycoplasma genitalium: a systematic review and meta-analysis. The Lancet Infectious diseases. 2020;20(11):1302-1314. Dixit A, Alexandrescu S, Boyer D, Graf EH, Vargas SO, Silverman M. Mycoplasma hominis Empyema in an 18-Year-old Stem Cell and Lung Transplant Recipient: Case Report and Review of the Literature. Journal of the Pediatric Infectious Diseases Society. 2017;6(4):e173-e176. Moneke I, Hornuss D, Serr A, Kern WV, Passlick B, Senbaklavaci O. Lung Abscess and Recurrent Empyema After Infection With Mycoplasma hominis: A Case Report and Review of the Literature. Open forum infectious diseases. 2022;9(1):ofab406. Wang Q, Tang X, van der Veen S. Mycoplasma hominis bloodstream infection and persistent pneumonia in a neurosurgery patient: a case report. BMC infectious diseases. 2022;22(1):169. Ahmed J, Rawre J, Dhawan N, Khanna N, Dhawan B. Mycoplasma hominis: An under recognized pathogen. Indian journal of medical microbiology. 2021;39(1):88-97. Wildenbeest JG, Said I, Jaeger B, van Hest RM, van de Beek D, Pajkrt D. Neonate with Mycoplasma hominis meningoencephalitis given moxifloxacin. The Lancet Infectious diseases. 2016;16(11):e261-e266. Combaz-Söhnchen N, Kuhn A. A Systematic Review of Mycoplasma and Ureaplasma in Urogynaecology. Geburtshilfe und Frauenheilkunde. 2017;77(12):1299-1303. Jin H, Xu X, Huang C, et al. Genomic characterisation of a multidrug-resistant Mycoplasma hominis isolate recovered from a synovial fluid sample in China. Journal of global antimicrobial resistance. 2020;20:282-284. Minion FC, Jarvill-Taylor K. Membrane-associated hemolysin activities in mycoplasmas. FEMS microbiology letters. 1994;116(1):101-106. Wu Y, Majidzadeh N, Li Y, et al. Trends of fluoroquinolones resistance in Mycoplasma and Ureaplasma urogenital isolates: Systematic review and meta-analysis. Journal of global antimicrobial resistance. 2024;36:13-25. Krausse R, Schubert S. In-vitro activities of tetracyclines, macrolides, fluoroquinolones and clindamycin against Mycoplasma hominis and Ureaplasma ssp. isolated in Germany over 20 years. Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases. 2010;16(11):1649-1655. Table Table 1. Antimicrobial susceptibility testing for M. hominis strain ZY_MH01. MIC (μg/mL) Breakpoints Interpretation Doxycycline 4 S≤4, R≥8 S Ofloxacin 8 S≤1, R≥4 R Erythromycin 8 S≤1, R≥4 R Tetracycline 4 S≤4, R≥8 S Clarithromycin 8 S≤1, R≥4 R Pristinamycin 2 R≥2 R Josamycin 2 S≤2, R≥8 S Ciprofloxacin 4 S≤1, R≥2 R Azithromycin 8 S≤0.12, R≥4 R the breakpoints were determined based on the guideline M43-A of Methods for antimicrobial susceptibility testing for human mycoplasmas approved by the Clinical and Laboratory Standards Institute. Additional Declarations No competing interests reported. Supplementary Files supplementaltable.1.xlsx Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: Revision requested 04 May, 2026 Reviews received at journal 30 Apr, 2026 Reviews received at journal 27 Apr, 2026 Reviews received at journal 12 Apr, 2026 Reviewers agreed at journal 10 Apr, 2026 Reviewers agreed at journal 07 Apr, 2026 Reviewers agreed at journal 07 Apr, 2026 Reviewers agreed at journal 04 Apr, 2026 Reviewers invited by journal 04 Apr, 2026 Editor assigned by journal 23 Mar, 2026 Submission checks completed at journal 21 Mar, 2026 First submitted to journal 21 Mar, 2026 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. 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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-9183112","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":621708876,"identity":"cbc57f75-0e4b-4453-9656-5622e03c6b8b","order_by":0,"name":"Hangfei Chen","email":"","orcid":"","institution":"Zhejiang University","correspondingAuthor":false,"prefix":"","firstName":"Hangfei","middleName":"","lastName":"Chen","suffix":""},{"id":621708878,"identity":"e445fd95-be5b-4a42-af8d-bd700088250f","order_by":1,"name":"Zhenping Wu","email":"","orcid":"","institution":"Zhejiang University","correspondingAuthor":false,"prefix":"","firstName":"Zhenping","middleName":"","lastName":"Wu","suffix":""},{"id":621708879,"identity":"49acf34e-ae91-451f-adab-7e7d2f6acc33","order_by":2,"name":"Qing Yang","email":"","orcid":"","institution":"Zhejiang University","correspondingAuthor":false,"prefix":"","firstName":"Qing","middleName":"","lastName":"Yang","suffix":""},{"id":621708881,"identity":"04717664-44eb-4888-a56a-8e59be14ef43","order_by":3,"name":"Wenqiao Yu","email":"","orcid":"","institution":"Zhejiang University","correspondingAuthor":false,"prefix":"","firstName":"Wenqiao","middleName":"","lastName":"Yu","suffix":""},{"id":621708882,"identity":"b46b989b-e66d-4d7d-b2d7-531d347c3a8e","order_by":4,"name":"Hua Zhou","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA2ElEQVRIiWNgGAWjYDACCQYGZiDFw8/AkMAAYROpRUaygVQtNgYHwFwitMjPbj4mXdhmx2N8/sAzCYYK68QG9rMH8GphnHMsTXpmWzKP2YEDaRIMZ9ITG3jyEvBqYZbIMZPmbWPmMTvYkCbB2HY4sUGCxwCvFjaIlnoe42YGoJZ/RGjhgWg5zGPABtLSQIQWCYm0ZGuec8d5JM4wJFskHEs3buPJwa9Ffkbywds8ZdX2/P1nEm98qLGW7Wc/g18LshsTwJHJRqx6IGA/QILiUTAKRsEoGEkAAKpsOUY/2vGLAAAAAElFTkSuQmCC","orcid":"","institution":"Zhejiang University","correspondingAuthor":true,"prefix":"","firstName":"Hua","middleName":"","lastName":"Zhou","suffix":""}],"badges":[],"createdAt":"2026-03-21 04:38:15","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-9183112/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9183112/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":106960215,"identity":"e557ebe8-9931-442d-a634-9c03ec6075dd","added_by":"auto","created_at":"2026-04-15 09:19:24","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":402137,"visible":true,"origin":"","legend":"\u003cp\u003eCT scan lung window (A) and mediastinum window (B) performed on April 6\u003csup\u003eth\u003c/sup\u003e 2024 (on the day of admission), presented with symptoms of chest infection and shortness of breath. Lung window (C) and mediastinum window (D) of the repeat CT scan performed on May 3\u003csup\u003erd\u003c/sup\u003e 2024, after 3 weeks of omadacycline treatment.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-9183112/v1/d79ba27aa43517e03212c03a.png"},{"id":106960354,"identity":"6c8ea287-5863-479e-866c-b768a33af3a7","added_by":"auto","created_at":"2026-04-15 09:20:25","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":362704,"visible":true,"origin":"","legend":"\u003cp\u003eCompleted genome sequence of \u003cem\u003eM. hominis\u003c/em\u003e ZY_MH01. Each arrow represents a gene, with green arrows indicating RNA.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-9183112/v1/ac18a194125bed52f8d095a8.png"},{"id":106835634,"identity":"0692b522-4a63-49c1-93e0-df53c5935392","added_by":"auto","created_at":"2026-04-14 02:01:47","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":209637,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic relationship between MH-1 and 50 other \u003cem\u003eM. hominis\u003c/em\u003e strains currently available in the NCBI GenBank database based on cgSNP analysis. Branch lengths were labled on the tree branches. The country where the strain was collected is indicated by the colored boxes, and the antibiotic resistance genes are indicated by the colored outermost triangles. The details of these strains (including organism infraspecific names and assembly accession numbers) can be found in Supplemental Table 1.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-9183112/v1/07c79df26502a8d933b770ae.png"},{"id":106962993,"identity":"660fd7d5-655a-4787-bbd5-f46049cb77ad","added_by":"auto","created_at":"2026-04-15 09:41:36","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1267467,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9183112/v1/0e0e301e-cc39-4e52-b713-468775b97346.pdf"},{"id":106835632,"identity":"b3c8bf0d-6567-4133-ba2d-6bbbc83cf8df","added_by":"auto","created_at":"2026-04-14 02:01:47","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":12505,"visible":true,"origin":"","legend":"","description":"","filename":"supplementaltable.1.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-9183112/v1/c8d83c8ab666d9fbf0e5d083.xlsx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Genomic Insights and Recent Epidemiology of a Multidrug-Resistant Mycoplasma hominis Isolate in China: Implications for Clinical Management","fulltext":[{"header":"Introduction","content":"\u003cp\u003e \u003cem\u003eM. hominis\u003c/em\u003e belongs to the genus Mycoplasma, which is commonly classified into two categories: genital and respiratory Mycoplasmas. The former category encompasses \u003cem\u003eM. hominis\u003c/em\u003e and \u003cem\u003eUreaplasma urealyticum\u003c/em\u003e, as well as \u003cem\u003eUreaplasma parvum\u003c/em\u003e, all of which are frequently detected in the human urogenital tract in both symptomatic patients and healthy individuals. However, apart from a few cases that lead to genital tract infections and neonatal infections\u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e, most individuals do not develop any disease\u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. In recent decades, multiple cases of extra-genital infections have been reported, including postoperative site infections such as osteitis and arthritis, mediastinitis, endocarditis, peritonitis, pyelonephritis, brain abscess, pneumonia, and wound infections, mainly in immunosuppressed patients. In addition, similar cases have also been described in immunocompetent patients, particularly in individuals with predisposing factors such as surgery and trauma. Respiratory infection with \u003cem\u003eM. hominis\u003c/em\u003e is relatively rare and primarily occurs in the form of disseminated infection in immunocompromised patients or patients with altered cardiorespiratory function\u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e~\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. In this study, the \u003cem\u003eM. hominis\u003c/em\u003e strain ZY_MH01 was collected from an immunocompetent patient with respiratory \u003cem\u003eM. hominis\u003c/em\u003e infection after blunt chest trauma. Additionally, the case's clinical features, imaging characteristics, and therapeutic options were analyzed by reviewing domestic and international literature.\u003c/p\u003e \u003cp\u003eA 47-year-old male fell from a height of eight meters and suffered bilateral multiple rib fractures and traumatic hemopneumothorax. The patient was previously healthy and his past medical history was insignificant. Chest Computed Tomography (CT) examination exhibited pelvic fractures, bilateral multiple rib fractures, and slight pleural effusion accompanied by atelectasis. The patient received thoracic cavity closed drainage and was managed conservatively. Fever and dyspnea appeared in six days. A repeat CT scan revealed more pleural effusion and slight pneumothorax. The patient developed respiratory failure and was intubated and placed under mechanical ventilation. The body temperature remained at 39.5℃; routine blood examination showed a white blood cell (WBC) count of 24.21*10\u003csup\u003e9\u003c/sup\u003e/L, a neutrophil count of 22.85*10\u003csup\u003e9\u003c/sup\u003e/L, C-reactive protein (CRP) levels of 146.73mg/L (normal value\u0026thinsp;\u0026lt;\u0026thinsp;10 mg/L), and procalcitonin levels of 4.75ng/mL. Empirical therapy was initiated for a suspected bacterial infection: cefoperazone sulbactam sodium (2.0g ivgtt q8h) combined with vancomycin (1g ivgtt q12h) was given with expectorant and other supportive treatment. An additional chest drainage was performed and 200 mL of light red pleural effusion was removed. The patient's body temperature and pulse rate temporarily decreased to 37.5℃ and 63/min, respectively. Subsequently, both parameters increased and remained elevated. A repeat chest CT scan showed patchy and increased density shadows, suggesting the possibility of a rapidly progressing lung infection (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA, B). The results of pleural effusion culture revealed pin-point-sized colonies on Columbia blood agar plates, which were identified as \u003cem\u003eM. hominis\u003c/em\u003e by MALDI-TOF MS. Antimicrobial treatment was switched to omadacycline (0.1g ivgtt qd), and the antimicrobial susceptibility testing was determined by VITEK 2 Compact System, demonstrating that the \u003cem\u003eM. hominis\u003c/em\u003e strain was resistant to clarithromycin and ciprofloxacin, but sensitive to tetracyclic (Table.1). The patient\u0026rsquo;s general condition, CRP, and oxygenation gradually returned to normal in the following 7 days, and omadacycline was stopped after total treatment duration of 18 days. The repeat chest CT showed that the majority of niduses returned to normal, with only minor pleural effusion (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eC, D). The patient fully recovered and was discharged a few days later.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"Methods and Results","content":"\u003cp\u003eThe \u003cem\u003eM. hominis\u003c/em\u003e strain ZY_MH01 was further confirmed by WGS. Total genomic DNA was extracted using a QIAamp DNA Mini Kit (Qiagen, Valencia, CA, USA) according to the manufacturer\u0026rsquo;s instructions. The quality of the extracted DNA was examined by a NanoDrop spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). A DNA library was then prepared using a Nextera DNA Library Preparation Kit (Illumina, San Diego, CA, USA), and WGS was performed on the Illumina HiSeq X10 platform (Illumina, San Diego, CA, USA) and long read Oxford Nanopore platform (Oxford, UK). After sequencing, all sequence reads were pre-processed to remove low-quality or artefactual bases. FastQC v.0.11.8 and Filtlong were separately used to assess the quality of the shotgun and long-read raw data. The clean data have been hybrid assembled by Unicycler.\u0026nbsp;The Whole Genome project has been deposited at DDBJ/ENA/GenBank under the accession JBFWID000000000.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFurthermore, the completed genome sequence of \u003cem\u003eM. hominis\u003c/em\u003e ZY_MH01 was subsequently assigned using Prokka v1.2.0. The sequence comprises 686,552 bp, with the overall G+C content accounting for 27.1% (Figure 2). Gene prediction was performed using Prodigal in single-genome mode to avoid overprediction commonly observed in small complete bacterial genomes. In total, 604 coding sequences, including 1 tmRNA, 4 rRNA and 33 tRNA genes, were identified. Virulence factors and antibiotic resistance were identified by the online web servers Virulence Factors Database (VFDB)-2022 and Comprehensive Antibiotic Resistance Database (CARD)-2024, which identified that the strain ZY_MH01 contains virulence genes \u003cem\u003etuf\u003c/em\u003e (elongation factor) and\u003cem\u003e\u0026nbsp;hlyA\u003c/em\u003e (membrane-associated hemolysin) and antimicrobial resistance mutation in gene\u003cem\u003e\u0026nbsp;parC\u0026nbsp;\u003c/em\u003e(S91I) (fluoroquinolone resistant) compare to the reference genome\u003cem\u003e\u0026nbsp;M. hominis\u003c/em\u003e ATCC 23114 (GenBank accession no. FP236530.1).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe phylogenetic relationship between \u003cem\u003eM. hominis\u003c/em\u003e ZY_MH01 and 50 \u003cem\u003eM. hominis\u003c/em\u003e strains with comparable sequence assembly levels, currently available in the NCBI GenBank database, was analyzed (Figure 3). These strains were recovered from Russia (21 strains), the UK (20 strains), the USA (5 strains), China (3 strains), and France (1 strain), respectively. The data suggested that these strains were epidemiologically related, and the antimicrobial resistance genes also exhibited regional associations. The closest relative of ZY_MH01 is MH-1, which was also recovered from Hangzhou, China, and this strain shares the same antimicrobial resistance mutation \u003cem\u003eparC\u0026nbsp;\u003c/em\u003e(S91I) and virulence genes \u003cem\u003etuf\u003c/em\u003e and\u003cem\u003e\u0026nbsp;hlyA\u003c/em\u003e.\u003c/p\u003e"},{"header":"Discussion and Conclusion","content":"\u003cp\u003e\u003cem\u003eM. hominis\u003c/em\u003e belongs to the genus \u003cem\u003eMycoplasma\u003c/em\u003e of the \u003cem\u003eMycoplasmataceae\u003c/em\u003e family within the \u003cem\u003eMollicutes\u003c/em\u003e class,\u0026nbsp;which\u0026nbsp;is characterized by the absence of a cell wall.\u0026nbsp;\u003cem\u003eM. hominis\u003c/em\u003e is a fastidious slow-growing organism that is hard to detect with routine laboratory methods, including gram staining and\u0026nbsp;routine bacterial culture\u003csup\u003e6,7\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThis study\u0026nbsp;reports a \u003cem\u003eM. hominis\u003c/em\u003e strain\u0026nbsp;ZY_MH01 collected\u0026nbsp;from a\u0026nbsp;patient\u0026nbsp;with\u0026nbsp;pleuropneumonia. Due to the characteristic\u0026nbsp;lack of a rigid cell wall, ZY_MH01\u0026nbsp;is intrinsically resistant to antimicrobials targeting the cell wall, including β-lactam antibiotics and\u0026nbsp;vancomycin, leading to empirical therapy failure.\u0026nbsp;Timely positive bacterial culture results promote quick treatment of the infection. However, \u003cem\u003eM. hominis\u003c/em\u003e is difficult to diagnose due to the microbe’s sensitivity to environmental factors and high demands for living conditions, thereby requiring delicate culture methods to yield a positive result\u003csup\u003e8\u003c/sup\u003e.\u0026nbsp;Conventional microbiological tests, such as\u0026nbsp;culture, smear microscopy, and PCR, require prior assumptions. Nonetheless, combining conventional tests with next-generation sequencing, including untargeted metagenomic or targeted approaches, could enhance pathogen diagnosis and further identify epidemiological information, virulence, and resistance genes.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe phylogenetic analysis revealed that \u003cem\u003eM. hominis\u003c/em\u003e demonstrated epidemiological relatedness, a factor previously deemed irrelevant in earlier research\u003csup\u003e9\u003c/sup\u003e, potentially due to the limited number of reported strains. Additionally, the closest phylogenetic relatives were found to frequently possess identical antimicrobial resistance genes and virulence genes. Both ZY_MH01 and its closest relative MH-1 contain the virulence genes \u003cem\u003etuf\u003c/em\u003e and\u003cem\u003e\u0026nbsp;hlyA\u003c/em\u003e, which indicate that their pathogenicity is similar to common strains of \u003cem\u003eMycoplasma\u0026nbsp;\u003c/em\u003espp\u003csup\u003e10\u003c/sup\u003e. The fluoroquinolone resistance gene \u003cem\u003eparC\u003c/em\u003e has been identified in ofloxacin and ciprofloxacin-resistant phenotypes, which is in accordance with a recent meta-analysis, revealing an increase of 59.8% and 31.2% in the resistance rate of ciprofloxacin and ofloxacin, respectively\u003csup\u003e11\u003c/sup\u003e. Moreover, in contrast to other \u003cem\u003eMycoplasma\u003c/em\u003e spp., \u003cem\u003eM. hominis\u003c/em\u003e is intrinsically resistant to macrolide antibiotics; hence tetracyclines are recommended as the first-line agents for mycoplasma\u003csup\u003e2,12\u003c/sup\u003e.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn summary, \u003cem\u003eM. hominis\u003c/em\u003e is not typically considered a common etiological agent of pleuropneumonia, and its identification poses challenges due to the necessity for extended anaerobic culturing. Molecular diagnostic techniques are essential and serve as a critical adjunct to traditional culture methods. Clinicians should remain cognizant of the potential involvement of this organism in a range of infections, particularly in immunocompromised individuals or those with a history of surgical procedures, trauma, or invasive catheterization. These findings enhance the understanding of the pathogen's genomic epidemiology and resistance mechanisms, thereby aiding timely diagnosis and appropriate antibiotic treatment to prevent long-term complications.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cdiv class=\"DefinitionList\"\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003e\u003cem\u003eM. hominis\u003c/em\u003e\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003e \u003cem\u003eMycoplasma hominis\u003c/em\u003e \u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eMALDI-TOF MS\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ematrix-assisted laser desorption/ionization time-of-flight mass spectrometry\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eCT\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eComputed tomography\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eCRP\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eC-reactive protein\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eWBC\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003ewhite blood cell\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv class=\"DefinitionListEntry\"\u003e \u003cdiv class=\"Term\"\u003eMIC\u003c/div\u003e \u003cdiv class=\"Description\"\u003e \u003cp\u003eminimum inhibiting concentration\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eHua Zhou and Wenqiao Yu were responsible for the organization and coordination of the study. Hangfei Chen was responsible for the data analysis and the finalization of the manuscript. Zhenping Wu served as the chief investigator, and Qtng Yang provided diagnostic assistance. All authors contributed to the writing of the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent and research ethics\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study has been reviewed and approved by the Research Ethics Committee of the First Affiliated Hospital of Zhejiang University (ITT20240876). The patient provided informed consent for publication of the clinical details including lung CT images, and written informed consent was obtained.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDisclosure\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no conflicts of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by research grants, including the Key R\u0026amp;D Plan of the Ministry of Science and Technology of China [grant number 2022YFC2504502]; the Research and Development Program of Zhejiang Province [grant number 2023C03068]; the National Natural Science Foundation of China [grant number 82272338], awarded to Hua Zhou. The funders were not involved in the study design, data collection and analysis, the decision to publish, or the preparation of the manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eCapoccia R, Greub G, Baud D. Ureaplasma urealyticum, Mycoplasma hominis and adverse pregnancy outcomes. \u003cem\u003eCurrent opinion in infectious diseases.\u0026nbsp;\u003c/em\u003e2013;26(3):231-240.\u003c/li\u003e\n \u003cli\u003eMachalek DA, Tao Y, Shilling H, et al. Prevalence of mutations associated with resistance to macrolides and fluoroquinolones in Mycoplasma genitalium: a systematic review and meta-analysis. \u003cem\u003eThe Lancet Infectious diseases.\u0026nbsp;\u003c/em\u003e2020;20(11):1302-1314.\u003c/li\u003e\n \u003cli\u003eDixit A, Alexandrescu S, Boyer D, Graf EH, Vargas SO, Silverman M. Mycoplasma hominis Empyema in an 18-Year-old Stem Cell and Lung Transplant Recipient: Case Report and Review of the Literature. \u003cem\u003eJournal of the Pediatric Infectious Diseases Society.\u0026nbsp;\u003c/em\u003e2017;6(4):e173-e176.\u003c/li\u003e\n \u003cli\u003eMoneke I, Hornuss D, Serr A, Kern WV, Passlick B, Senbaklavaci O. Lung Abscess and Recurrent Empyema After Infection With Mycoplasma hominis: A Case Report and Review of the Literature. \u003cem\u003eOpen forum infectious diseases.\u0026nbsp;\u003c/em\u003e2022;9(1):ofab406.\u003c/li\u003e\n \u003cli\u003eWang Q, Tang X, van der Veen S. Mycoplasma hominis bloodstream infection and persistent pneumonia in a neurosurgery patient: a case report. \u003cem\u003eBMC infectious diseases.\u0026nbsp;\u003c/em\u003e2022;22(1):169.\u003c/li\u003e\n \u003cli\u003eAhmed J, Rawre J, Dhawan N, Khanna N, Dhawan B. Mycoplasma hominis: An under recognized pathogen. \u003cem\u003eIndian journal of medical microbiology.\u0026nbsp;\u003c/em\u003e2021;39(1):88-97.\u003c/li\u003e\n \u003cli\u003eWildenbeest JG, Said I, Jaeger B, van Hest RM, van de Beek D, Pajkrt D. Neonate with Mycoplasma hominis meningoencephalitis given moxifloxacin. \u003cem\u003eThe Lancet Infectious diseases.\u0026nbsp;\u003c/em\u003e2016;16(11):e261-e266.\u003c/li\u003e\n \u003cli\u003eCombaz-S\u0026ouml;hnchen N, Kuhn A. A Systematic Review of Mycoplasma and Ureaplasma in Urogynaecology. \u003cem\u003eGeburtshilfe und Frauenheilkunde.\u0026nbsp;\u003c/em\u003e2017;77(12):1299-1303.\u003c/li\u003e\n \u003cli\u003eJin H, Xu X, Huang C, et al. Genomic characterisation of a multidrug-resistant Mycoplasma hominis isolate recovered from a synovial fluid sample in China. \u003cem\u003eJournal of global antimicrobial resistance.\u0026nbsp;\u003c/em\u003e2020;20:282-284.\u003c/li\u003e\n \u003cli\u003eMinion FC, Jarvill-Taylor K. Membrane-associated hemolysin activities in mycoplasmas. \u003cem\u003eFEMS microbiology letters.\u0026nbsp;\u003c/em\u003e1994;116(1):101-106.\u003c/li\u003e\n \u003cli\u003eWu Y, Majidzadeh N, Li Y, et al. Trends of fluoroquinolones resistance in Mycoplasma and Ureaplasma urogenital isolates: Systematic review and meta-analysis. \u003cem\u003eJournal of global antimicrobial resistance.\u0026nbsp;\u003c/em\u003e2024;36:13-25.\u003c/li\u003e\n \u003cli\u003eKrausse R, Schubert S. In-vitro activities of tetracyclines, macrolides, fluoroquinolones and clindamycin against Mycoplasma hominis and Ureaplasma ssp. isolated in Germany over 20 years. \u003cem\u003eClinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.\u0026nbsp;\u003c/em\u003e2010;16(11):1649-1655.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Table","content":"\u003cp\u003eTable 1. Antimicrobial susceptibility testing for \u003cem\u003eM. hominis\u003c/em\u003e strain ZY_MH01.\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\" width=\"550\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd nowrap=\"\" style=\"width: 118px;\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 153px;\"\u003e\n \u003cp\u003eMIC (\u0026mu;g/mL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 138px;\"\u003e\n \u003cp\u003eBreakpoints\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003eInterpretation\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd nowrap=\"\" style=\"width: 118px;\"\u003e\n \u003cp\u003eDoxycycline\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 153px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 138px;\"\u003e\n \u003cp\u003eS\u0026le;4, R\u0026ge;8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd nowrap=\"\" style=\"width: 118px;\"\u003e\n \u003cp\u003eOfloxacin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 153px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 138px;\"\u003e\n \u003cp\u003eS\u0026le;1, R\u0026ge;4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd nowrap=\"\" style=\"width: 118px;\"\u003e\n \u003cp\u003eErythromycin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 153px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 138px;\"\u003e\n \u003cp\u003eS\u0026le;1, R\u0026ge;4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd nowrap=\"\" style=\"width: 118px;\"\u003e\n \u003cp\u003eTetracycline\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 153px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 138px;\"\u003e\n \u003cp\u003eS\u0026le;4, R\u0026ge;8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd nowrap=\"\" style=\"width: 118px;\"\u003e\n \u003cp\u003eClarithromycin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 153px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 138px;\"\u003e\n \u003cp\u003eS\u0026le;1, R\u0026ge;4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd nowrap=\"\" style=\"width: 118px;\"\u003e\n \u003cp\u003ePristinamycin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 153px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 138px;\"\u003e\n \u003cp\u003eR\u0026ge;2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd nowrap=\"\" style=\"width: 118px;\"\u003e\n \u003cp\u003eJosamycin\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 153px;\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 138px;\"\u003e\n \u003cp\u003eS\u0026le;2, R\u0026ge;8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003eS\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd nowrap=\"\" style=\"width: 118px;\"\u003e\n \u003cp\u003eCiprofloxacin\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 153px;\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 138px;\"\u003e\n \u003cp\u003eS\u0026le;1, R\u0026ge;2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd nowrap=\"\" style=\"width: 118px;\"\u003e\n \u003cp\u003eAzithromycin\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 153px;\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 138px;\"\u003e\n \u003cp\u003eS\u0026le;0.12, R\u0026ge;4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 141px;\"\u003e\n \u003cp\u003eR\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003ethe breakpoints were determined based on the guideline M43-A of Methods for antimicrobial susceptibility testing for human mycoplasmas approved by the Clinical and Laboratory Standards Institute.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":false,"email":"","identity":"current-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Current Microbiology","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"VoR Journals","inReviewEnabled":false,"inReviewRevisionsEnabled":false},"keywords":"Mycoplasma hominis, pneumonia, immunocompetent, multidrug-resistance, cgSNP","lastPublishedDoi":"10.21203/rs.3.rs-9183112/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9183112/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cem\u003eMycoplasma hominis (M. hominis) is an opportunistic pathogen linked to urogenital and neonatal infections; however, limited genetic and epidemiological data are available. Extragenital infections in healthy adults are rare, and effective antibiotics are species-specific, complicating diagnosis and treatment. Hence, this study aimed to elucidate the clinical process, update the epidemiological characteristics, and investigate the genomic features of a multidrug-resistant M. hominis isolate from an immunocompetent pleuropneumonia patient in China. \u003c/em\u003eThe\u003cem\u003e M. hominis\u003c/em\u003e isolate ZY_MH01 was recovered from pleural fluid and was identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and Whole Genome Sequencing (WGS). The completed genome was annotated using the NCBI Prokaryotic Genome Annotation Pipeline (PGAP). Snippy v4.4.5 was utilized to conduct a core genome single nucleotide polymorphism (cgSNP) analysis between ZY_MH01 and 50 \u003cem\u003eM. hominis\u003c/em\u003e strains from the NCBI GenBank database. Subsequently, phylogenies were constructed using IQtree v2.0.3 and visualized by iTOL. Antimicrobial resistance genes and susceptibility were identified by CARD RGI v6.0.3 and VITEK 2 Compact System. The antimicrobial resistance mutation in gene \u003cem\u003eparC\u003c/em\u003e (S91I) \u0026nbsp;was found in the genome. Phylogenetic analysis revealed that ZY_MH01 and strains from the NCBI database were epidemiologically related, with ZY_MH01's closest relatives being in the same city and sharing the same resistance and virulence genes. \u003cem\u003eM. hominis\u003c/em\u003e should be considered a potential cause of pulmonary infection, particularly in patients unresponsive to broad-spectrum antibiotics. This study provides a detailed analysis of \u003cem\u003eM. hominis\u003c/em\u003e epidemiology, highlighting regional evolutionary relationships among strains.\u003c/p\u003e","manuscriptTitle":"Genomic Insights and Recent Epidemiology of a Multidrug-Resistant Mycoplasma hominis Isolate in China: Implications for Clinical Management","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-14 02:01:43","doi":"10.21203/rs.3.rs-9183112/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-05-04T04:22:02+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-30T15:28:03+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-27T09:55:00+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-12T14:06:58+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"101424737776509827460663662714596568908","date":"2026-04-10T12:43:27+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"65024098200397553101756840693304429922","date":"2026-04-07T19:45:54+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"329299372342420173348224601093660630301","date":"2026-04-07T13:45:06+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"276025690339344717124099098960456325542","date":"2026-04-04T19:16:10+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-04-04T18:29:28+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-03-23T12:05:20+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-03-22T00:55:15+00:00","index":"","fulltext":""},{"type":"submitted","content":"Current Microbiology","date":"2026-03-21T04:22:22+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":false,"email":"","identity":"current-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Current Microbiology","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"VoR Journals","inReviewEnabled":false,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"8657725b-f7c0-4806-b001-b1caf4ce4f2f","owner":[],"postedDate":"April 14th, 2026","published":true,"recentEditorialEvents":[{"type":"decision","content":"Revision requested","date":"2026-05-04T04:22:02+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-30T15:28:03+00:00","index":23,"fulltext":""}],"rejectedJournal":[],"revision":"","amendment":"","status":"in-revision","subjectAreas":[],"tags":[],"updatedAt":"2026-05-04T04:24:57+00:00","versionOfRecord":[],"versionCreatedAt":"2026-04-14 02:01:43","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9183112","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9183112","identity":"rs-9183112","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}