Molecular Assessment of Bacterial Vaginosis by Lactobacillus Abundance and Species Diversity

Authors

Joke A. M. Dols, VU University Amsterdam
Douwe Molenaar, VU University Amsterdam
Jannie J. van der Helm, Public Health Service of Amsterdam
Martien P. M. Caspers, Netherlands Organisation for Applied Scientific Research
Alie de Kat Angelino-Bart, Netherlands Organisation for Applied Scientific Research
Frank H. J. Schuren, Netherlands Organisation for Applied Scientific Research
Adrianus G. C. L. Speksnijder, Public Health Service of Amsterdam
Hans V. Westerhoff, VU University Amsterdam
Jan Hendrik Richardus, University Medical Center Rotterdam
Mathilde E. Boon, Leiden Cytology and Pathology Laboratory
Gregor Reid, Lawson Health Research Institute; Western UniversityFollow
Henry J. C. de Vries, Public Health Service of Amsterdam
Remco Kort, VU University Amsterdam

Document Type

Article

Publication Date

4-2016

Journal

BMC Infectious Diseases

Volume

16

URL with Digital Object Identifier

http://dx.doi.org/10.1186/s12879-016-1513-3

Abstract

Background

To date, women are most often diagnosed with bacterial vaginosis (BV) using microscopy based Nugent scoring or Amsel criteria. However, the accuracy is less than optimal. The aim of the present study was to confirm the identity of known BV-associated composition profiles and evaluate indicators for BV using three molecular methods.

Methods

Evaluation of indicators for BV was carried out by 16S rRNA amplicon sequencing of the V5-V7 region, a tailor-made 16S rRNA oligonucleotide-based microarray, and a PCR-based profiling technique termed IS-profiling, which is based on fragment variability of the 16S-23S rRNA intergenic spacer region. An inventory of vaginal bacterial species was obtained from 40 females attending a Dutch sexually transmitted infection outpatient clinic, of which 20 diagnosed with BV (Nugent score 7–10), and 20 BV negative (Nugent score 0–3).

Results

Analysis of the bacterial communities by 16S rRNA amplicon sequencing revealed two clusters in the BV negative women, dominated by either Lactobacillus iners or Lactobacillus crispatus and three distinct clusters in the BV positive women. In the former, there was a virtually complete, negative correlation between L. crispatus and L. iners. BV positive subjects showed cluster profiles that were relatively high in bacterial species diversity and dominated by anaerobic species, including Gardnerella vaginalis, and those belonging to the Families of Lachnospiraceae and Leptotrichiaceae. Accordingly, the Gini-Simpson index of species diversity, and the relative abundance Lactobacillus species appeared consistent indicators for BV. Under the conditions used, only the 16S rRNA amplicon sequencing method was suitable to assess species diversity, while all three molecular composition profiling methods were able to indicate Lactobacillus abundance in the vaginal microbiota.

Conclusion

An affordable and simple molecular test showing a depletion of the genus Lactobacillus in combination with an increased species diversity of vaginal microbiota could serve as an alternative and practical diagnostic method for the assessment of BV.

Notes

e-Publication Number: 180

Western Libraries Open Access Fund recipient.