Development and analytical validation of real-time PCR for the detection of Streptococcus agalactiae in pregnant women

Daniel F. Escobar, Diego A. Diaz-Dinamarca, Carlos F. Hernández, Daniel A. Soto, Ricardo A. Manzo, Pedro I. Alarcón, Camila H. Pinto, Diego N. Bastias, Carolayn N. Oberg-Bravo, Robert Rojas, Sebastián E. Illanes, Alexis M. Kalergis, Abel E. Vasquez

Research output: Contribution to journalArticlepeer-review

Abstract

BACKGROUND: Group B Streptococcus (GBS) is the leading cause of invasive neonatal infection. In this study, we aimed to evaluate the analytical validation of qualitative real-time polymerase chain reaction (qPCR) as a means to detect GBS. METHODS: Genomic DNA (gDNA) was purified from 12 ATCC bacterial strains, two belonging to GBS and the remainder acting as negative controls. Additionally, gDNA was isolated from 21 strains of GBS from various serotypes (Ia, Ib and II-VIII). All gDNA was used to evaluate the analytical validation of the qPCR method employing a specific Taqman probe. Inclusivity, exclusivity, anticipated reportable range, the limit of detection and robustness were evaluated. The methods used are described in international guidelines and other existing reports. The performance of this qPCR method for detecting GBS was compared to other microbiological methods used with vaginal-rectal samples from pregnant women. RESULTS: Our qPCR method for detecting GBS was analytically validated. It has a limit of detection of 0.7 GE/μL and 100% analytical specificity. It detects all strains of GBS with the same level of performance as microbiological methods. CONCLUSION: Data suggest that this qPCR method performs adequately as a means to detect GBS in vaginal-rectal swabs from pregnant women.
Original languageAmerican English
Pages (from-to)352
Number of pages1
JournalBMC Pregnancy and Childbirth
Volume20
Issue number1
DOIs
StatePublished - 9 Jun 2020

Keywords

  • Analytical validation
  • Bacterial detection
  • Group B Streptococcus
  • qPCR
  • Surface immunogenic protein

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