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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.

Evaluation of a novel real-time PCR test based on the ssrAgene for the identification of group B streptococci in vaginal swabs