Dan Duy Pham-Tran, Lex E X Leong, Mark McMillan, Andrew Lawrence, Hassen Mohammed, Adam Finn, Helen S Marshall
{"title":"基于sodc的PCR检测青少年咽拭子携带脑膜炎奈瑟菌缺乏特异性。","authors":"Dan Duy Pham-Tran, Lex E X Leong, Mark McMillan, Andrew Lawrence, Hassen Mohammed, Adam Finn, Helen S Marshall","doi":"10.1128/spectrum.02779-24","DOIUrl":null,"url":null,"abstract":"<p><p>The utility of the superoxide dismutase C (<i>sodC</i>) gene for detecting <i>Neisseria meningitidis</i> carriage in pharyngeal swab samples is unclear. While it has been explored as an alternative to the capsule transport A gene for detecting invasive strains, its potential as the sole target for real-time PCR in meningococcal carriage screening remains unexplored. This study aimed to assess the utility of using <i>sodC</i> as a target for detecting invasive and non-invasive strains of <i>Neisseria meningitidis</i> in pharyngeal swabs. Oropharyngeal swab samples collected from adolescents (approximately 15-18 years of age) enrolled in the \"B-Part-of-It\" cluster-randomized controlled trial (NCT03089086) were selected for this study. Samples were initially screened for the presence of specific <i>N meningitidis</i> porin A gene (<i>porA</i>) using PCR and then preserved by freezing at -80°C. An assay targeting the <i>sodC</i> gene was developed using the locked nucleic acid probe-based method. Upon optimization of the <i>sodC</i> assay, 1,092 samples were retested using this assay and compared against the results from the <i>porA</i> assays to determine concordance. Out of 1,092 samples tested, 965 (88.4%) were <i>sodC</i> positive, with 100% sensitivity and 13% specificity compared to <i>porA</i>. The positive predictive values and negative predictive values for the <i>sod</i>C assay were 12% and 100%, respectively. When compared to <i>porA</i> PCR as the gold standard, the <i>sodC</i> assay lacks sufficient specificity to serve as a stand-alone screening assay for the detection of <i>Neisseria meningitidis</i> carriage in pharyngeal samples from adolescents.IMPORTANCEWhile the <i>sodC</i> assay successfully detects <i>N. meningitidis</i>, we identified a limitation in its specificity due to potential cross-reactivity with other organisms, including <i>Haemophilus</i> spp., which can result in false positives. This limitation highlights the need for careful interpretation of <i>Neisseria meningitidis</i> carriage results, especially in epidemiological studies.</p>","PeriodicalId":18670,"journal":{"name":"Microbiology spectrum","volume":" ","pages":"e0277924"},"PeriodicalIF":3.7000,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"The lack of specificity of <i>sodC</i>-based PCR for the detection of <i>Neisseria meningitidis</i> carriage in pharyngeal swabs from adolescents.\",\"authors\":\"Dan Duy Pham-Tran, Lex E X Leong, Mark McMillan, Andrew Lawrence, Hassen Mohammed, Adam Finn, Helen S Marshall\",\"doi\":\"10.1128/spectrum.02779-24\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>The utility of the superoxide dismutase C (<i>sodC</i>) gene for detecting <i>Neisseria meningitidis</i> carriage in pharyngeal swab samples is unclear. While it has been explored as an alternative to the capsule transport A gene for detecting invasive strains, its potential as the sole target for real-time PCR in meningococcal carriage screening remains unexplored. This study aimed to assess the utility of using <i>sodC</i> as a target for detecting invasive and non-invasive strains of <i>Neisseria meningitidis</i> in pharyngeal swabs. Oropharyngeal swab samples collected from adolescents (approximately 15-18 years of age) enrolled in the \\\"B-Part-of-It\\\" cluster-randomized controlled trial (NCT03089086) were selected for this study. Samples were initially screened for the presence of specific <i>N meningitidis</i> porin A gene (<i>porA</i>) using PCR and then preserved by freezing at -80°C. An assay targeting the <i>sodC</i> gene was developed using the locked nucleic acid probe-based method. Upon optimization of the <i>sodC</i> assay, 1,092 samples were retested using this assay and compared against the results from the <i>porA</i> assays to determine concordance. Out of 1,092 samples tested, 965 (88.4%) were <i>sodC</i> positive, with 100% sensitivity and 13% specificity compared to <i>porA</i>. The positive predictive values and negative predictive values for the <i>sod</i>C assay were 12% and 100%, respectively. When compared to <i>porA</i> PCR as the gold standard, the <i>sodC</i> assay lacks sufficient specificity to serve as a stand-alone screening assay for the detection of <i>Neisseria meningitidis</i> carriage in pharyngeal samples from adolescents.IMPORTANCEWhile the <i>sodC</i> assay successfully detects <i>N. meningitidis</i>, we identified a limitation in its specificity due to potential cross-reactivity with other organisms, including <i>Haemophilus</i> spp., which can result in false positives. This limitation highlights the need for careful interpretation of <i>Neisseria meningitidis</i> carriage results, especially in epidemiological studies.</p>\",\"PeriodicalId\":18670,\"journal\":{\"name\":\"Microbiology spectrum\",\"volume\":\" \",\"pages\":\"e0277924\"},\"PeriodicalIF\":3.7000,\"publicationDate\":\"2025-04-08\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Microbiology spectrum\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1128/spectrum.02779-24\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Microbiology spectrum","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1128/spectrum.02779-24","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
The lack of specificity of sodC-based PCR for the detection of Neisseria meningitidis carriage in pharyngeal swabs from adolescents.
The utility of the superoxide dismutase C (sodC) gene for detecting Neisseria meningitidis carriage in pharyngeal swab samples is unclear. While it has been explored as an alternative to the capsule transport A gene for detecting invasive strains, its potential as the sole target for real-time PCR in meningococcal carriage screening remains unexplored. This study aimed to assess the utility of using sodC as a target for detecting invasive and non-invasive strains of Neisseria meningitidis in pharyngeal swabs. Oropharyngeal swab samples collected from adolescents (approximately 15-18 years of age) enrolled in the "B-Part-of-It" cluster-randomized controlled trial (NCT03089086) were selected for this study. Samples were initially screened for the presence of specific N meningitidis porin A gene (porA) using PCR and then preserved by freezing at -80°C. An assay targeting the sodC gene was developed using the locked nucleic acid probe-based method. Upon optimization of the sodC assay, 1,092 samples were retested using this assay and compared against the results from the porA assays to determine concordance. Out of 1,092 samples tested, 965 (88.4%) were sodC positive, with 100% sensitivity and 13% specificity compared to porA. The positive predictive values and negative predictive values for the sodC assay were 12% and 100%, respectively. When compared to porA PCR as the gold standard, the sodC assay lacks sufficient specificity to serve as a stand-alone screening assay for the detection of Neisseria meningitidis carriage in pharyngeal samples from adolescents.IMPORTANCEWhile the sodC assay successfully detects N. meningitidis, we identified a limitation in its specificity due to potential cross-reactivity with other organisms, including Haemophilus spp., which can result in false positives. This limitation highlights the need for careful interpretation of Neisseria meningitidis carriage results, especially in epidemiological studies.
期刊介绍:
Microbiology Spectrum publishes commissioned review articles on topics in microbiology representing ten content areas: Archaea; Food Microbiology; Bacterial Genetics, Cell Biology, and Physiology; Clinical Microbiology; Environmental Microbiology and Ecology; Eukaryotic Microbes; Genomics, Computational, and Synthetic Microbiology; Immunology; Pathogenesis; and Virology. Reviews are interrelated, with each review linking to other related content. A large board of Microbiology Spectrum editors aids in the development of topics for potential reviews and in the identification of an editor, or editors, who shepherd each collection.