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Multiplex detection of bacteria associated with normal microbiota and with bacterial vaginosis in vaginal swabs using oligonucleotide-coupled fluorescent microspheres
Journal of Clinical Microbiology, 10/08/09
Dumonceaux TJ et al. – The authors designed a 9–plex Luminex array for characterizing the vaginal microbiota and applied it to the analysis of vaginal swabs from African and North American individuals. Using the presence of one or both of A. vaginae and G. vaginalis as the defining criterion for BV, the authors found that the method showed a high sensitivity and specificity for diagnosing BV compared to traditional microscopy.
Tim J Dumonceaux, 10/08/09
| Bacterial vaginosis (BV) is commonly diagnosed using a set of microscopic criteria (Nugent or Hay-Ison scale) that, while rapid, accurate, and inexpensive, can understate the complexity of the vaginal microbiota and does not enable the profiling of the bacterial species present in the swab. We designed a Luminex-based array for rapidly determining the species profile of a vaginal swab using chaperonin-60 (cpn60)-targeted oligonucleotide probes. Multiplex detection was facilitated using universal cpn60-targeted amplification primers in combination with highly target-specific hybridization probes. The method was specific and sensitive for diagnosing BV compared with microscopy, and furthermore the amplitude of the Luminex signal correlated to the number of genomes of each organism that was detected using quantitative PCR. These features enabled the longitudinal profiling of the vaginal microbiota in a number of individuals, revealing the dynamics of the population in each individual over time. |
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