Multiplex detection of bacteria associated with normal microbiota and with bacterial vaginosis in vaginal swabs using oligonucleotide-coupled fluorescent microspheres.
Dumonceaux, T.J., Schellenberg, J.J., Goleski, V., Hill, J.E., Jaoko, W., Kimani, J., Money, D., Ball, T.B., Plummer, F.A., and Severini, A. (2009). "Multiplex detection of bacteria associated with normal microbiota and with bacterial vaginosis in vaginal swabs using oligonucleotide-coupled fluorescent microspheres.", Journal of Clinical Microbiology, 47(12), pp. 4067-4077. doi : 10.1128/JCM.00112-09 Access to full text
Bacterial vaginosis (BV) is a recurrent condition that is associated with a range of negative outcomes, including the acquisition of HIV and other sexually transmitted diseases, preterm births, and pelvic inflammatory disease. In contrast to the Lactobacillus-dominated normal vaginal microbiota, BV is characterized by a lack of lactobacilli and an abundance of anaerobic and Gram-negative organisms, including Garderella vaginalis and Atopobium vaginae. The laboratory diagnosis of BV to date has relied upon criteria observed by microscopy of Gram-stained vaginal swabs. We describe a molecular-based method for easily determining the species profile within the vaginal microbiota based on the amplification of the chaperonin-60 genes of all bacteria present in the swab and hybridization of the amplicon to species-specific oligonucleotide coupled fluorescent beads that are identified by flow cytometry using a Luminex instrument. We 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, we found that the method showed a high sensitivity and specificity for diagnosing BV compared to traditional microscopy.
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