Limitations of 16S rRNA Gene Sequencing to Characterize Lactobacillus Species in the Upper Genital Tract.

Autor:	O'Callaghan JL; Faculty of Health, Centre for Immunology and Infection Control, Queensland University of Technology, Brisbane, QLD, Australia.; Faculty of Health, School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD, Australia., Willner D; Australian Centre for Ecogenomics, University of Queensland, St Lucia, QLD, Australia.; Department of Computer Science, College of William & Mary, Williamsburg, VA, United States., Buttini M; The Wesley Hospital, Auchenflower, QLD, Australia., Huygens F; Faculty of Health, Centre for Immunology and Infection Control, Queensland University of Technology, Brisbane, QLD, Australia.; Faculty of Health, School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD, Australia., Pelzer ES; Faculty of Health, Centre for Immunology and Infection Control, Queensland University of Technology, Brisbane, QLD, Australia.; Faculty of Health, School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD, Australia.
Jazyk:	angličtina
Zdroj:	Frontiers in cell and developmental biology [Front Cell Dev Biol] 2021 Jul 29; Vol. 9, pp. 641921. Date of Electronic Publication: 2021 Jul 29 (Print Publication: 2021).
DOI:	10.3389/fcell.2021.641921
Abstrakt:	The endometrial cavity is an upper genital tract site previously thought as sterile, however, advances in culture-independent, next-generation sequencing technology have revealed that this low-biomass site harbors a rich microbial community which includes multiple Lactobacillus species. These bacteria are considered to be the most abundant non-pathogenic genital tract commensals. Next-generation sequencing of the female lower genital tract has revealed significant variation amongst microbial community composition with respect to Lactobacillus sp. in samples collected from healthy women and women with urogenital conditions. The aim of this study was to evaluate our ability to characterize members of the genital tract microbial community to species-level taxonomy using variable regions of the 16S rRNA gene. Samples were interrogated for the presence of microbial DNA using next-generation sequencing technology that targets the V5-V8 regions of the 16S rRNA gene and compared to speciation using qPCR. We also performed re-analysis of published data using alternate variable regions of the 16S rRNA gene. In this analysis, we explore next-generation sequencing of clinical genital tract isolates as a method for high throughput identification to species-level of key Lactobacillus sp. Data revealed that characterization of genital tract taxa is hindered by a lack of a consensus protocol and 16S rRNA gene region target allowing comparison between studies. Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2021 O’Callaghan, Willner, Buttini, Huygens and Pelzer.)
Databáze:	MEDLINE
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