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

Hdl Handle:
http://hdl.handle.net/10147/94027
Title:
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, Martina; Mullen, 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.ie
Citation:
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 diseases
Issue Date:
2009
URI:
http://hdl.handle.net/10147/94027
DOI:
10.1186/1471-2334-9-148
PubMed ID:
19732424
Abstract:
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:
en
MeSH:
DNA, Bacterial; Female; Genes, Bacterial; Humans; Polymerase Chain Reaction; Predictive Value of Tests; Pregnancy; RNA, Bacterial; Sensitivity and Specificity; Streptococcal Infections; Streptococcus agalactiae; Vagina
ISSN:
1471-2334

Full metadata record

DC FieldValue Language
dc.contributor.authorWernecke, Martinaen
dc.contributor.authorMullen, Ciaraen
dc.contributor.authorSharma, Vimlaen
dc.contributor.authorMorrison, Johnen
dc.contributor.authorBarry, Thomasen
dc.contributor.authorMaher, Majellaen
dc.contributor.authorSmith, Terryen
dc.date.accessioned2010-03-10T12:00:01Z-
dc.date.available2010-03-10T12:00:01Z-
dc.date.issued2009-
dc.identifier.citationEvaluation 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.en
dc.identifier.issn1471-2334-
dc.identifier.pmid19732424-
dc.identifier.doi10.1186/1471-2334-9-148-
dc.identifier.urihttp://hdl.handle.net/10147/94027-
dc.description.abstractBACKGROUND: 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.-
dc.language.isoenen
dc.subject.meshDNA, Bacterial-
dc.subject.meshFemale-
dc.subject.meshGenes, Bacterial-
dc.subject.meshHumans-
dc.subject.meshPolymerase Chain Reaction-
dc.subject.meshPredictive Value of Tests-
dc.subject.meshPregnancy-
dc.subject.meshRNA, Bacterial-
dc.subject.meshSensitivity and Specificity-
dc.subject.meshStreptococcal Infections-
dc.subject.meshStreptococcus agalactiae-
dc.subject.meshVagina-
dc.titleEvaluation of a novel real-time PCR test based on the ssrA gene for the identification of group B streptococci in vaginal swabs.en
dc.contributor.departmentMolecular Diagnostics Research Group, National Centre for Biomedical Engineering Science, Galway, Ireland. martina.wernecke@nuigalway.ieen
dc.identifier.journalBMC infectious diseasesen

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