Evaluation of a novel real-time PCR test based on the ssrA gene for the identification of group B streptococci in vaginal swabs.
Authors
Wernecke, MartinaMullen, Ciara
Sharma, Vimla
Morrison, John
Barry, Thomas
Maher, Majella
Smith, Terry
Affiliation
Molecular Diagnostics Research Group, National Centre for Biomedical Engineering Science, Galway, Ireland. martina.wernecke@nuigalway.ieIssue Date
2009MeSH
DNA, BacterialFemale
Genes, Bacterial
Humans
Polymerase Chain Reaction
Predictive Value of Tests
Pregnancy
RNA, Bacterial
Sensitivity and Specificity
Streptococcal Infections
Streptococcus agalactiae
Vagina
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Evaluation of a novel real-time PCR test based on the ssrA gene for the identification of group B streptococci in vaginal swabs. 2009, 9:148 BMC Infect. Dis.Journal
BMC infectious diseasesDOI
10.1186/1471-2334-9-148PubMed ID
19732424Abstract
BACKGROUND: Despite the implementation of prevention guidelines, early-onset group B streptococci (GBS) disease remains a cause of neonatal morbidity and mortality worldwide. Strategies to identify women who are at risk of transmitting GBS to their infant and the administration of intrapartum antibiotics have greatly reduced the incidence of neonatal GBS disease. However, there is a requirement for a rapid diagnostic test for GBS that can be carried out in a labour ward setting especially for women whose GBS colonisation status is unknown at the time of delivery. We report the design and evaluation of a real-time PCR test (RiboSEQ GBS test) for the identification of GBS in vaginal swabs from pregnant women. METHODS: The qualitative real-time PCR RiboSEQ GBS test was designed based on the bacterial ssrA gene and incorporates a competitive internal standard control. The analytical sensitivity of the test was established using crude lysate extracted from serial dilutions of overnight GBS culture using the IDI Lysis kit. Specificity studies were performed using DNA prepared from a panel of GBS strains, related streptococci and other species found in the genital tract environment. The RiboSEQ GBS test was evaluated on 159 vaginal swabs from pregnant women and compared with the GeneOhm StrepB Assay and culture for the identification of GBS. RESULTS: The RiboSEQ GBS test is specific and has an analytical sensitivity of 1-10 cell equivalents. The RiboSEQ GBS test was 96.4% sensitive and 95.8% specific compared to "gold standard" culture for the identification of GBS in vaginal swabs from pregnant women. In this study, the RiboSEQ GBS test performed slightly better than the commercial BD GeneOhm StrepB Assay which gave a sensitivity of 94.6% and a specificity of 89.6% compared to culture. CONCLUSION: The RiboSEQ GBS test is a valuable method for the rapid, sensitive and specific detection of GBS in pregnant women. This study also validates the ssrA gene as a suitable and versatile target for nucleic acid-based diagnostic tests for bacterial pathogens.Language
enISSN
1471-2334ae974a485f413a2113503eed53cd6c53
10.1186/1471-2334-9-148
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