Comparative analysis of vaginal microbiota sampling using menstrual cups and high vaginal swabs in pregnant women living with HIV-1 infection

Frontiers in Cellular and Infection Microbiology

Cellular and Infection Microbiology

Ann Smith¹  David A. MacIntyre²  Yun S. Lee²  Julian R. Marchesi³  Robin Shattock⁴  Veronica G. Preda⁵  Graham P. Taylor⁶  Charlotte-Eve S. Short⁶  Rachael Quinlan⁶  Phillip R. Bennett⁷ 

[1] Faculty of Health and Applied Sciences, University West of England, Bristol, United Kingdom;March of Dimes Prematurity Research Centre, Division of the Institute of Reproductive and Developmental Biology, Department of Metabolism, Digestion, and Reproduction, Imperial College London, London, United Kingdom;March of Dimes Prematurity Research Centre, Division of the Institute of Reproductive and Developmental Biology, Department of Metabolism, Digestion, and Reproduction, Imperial College London, London, United Kingdom;Marchesi Laboratory, Department of Metabolism, Digestion, and Reproduction, Division of Digestive Disease, Faculty of Medicine, Imperial College London, London, United Kingdom;Section of Immunology of Infection, Department of Infectious Disease, Imperial College London, London, United Kingdom;Section of Virology, Department of Infectious Disease, Imperial College London, London, United Kingdom;St Mary’s Hospital, Imperial College Healthcare NHS Trust, London, United Kingdom;Section of Virology, Department of Infectious Disease, Imperial College London, London, United Kingdom;St Mary’s Hospital, Imperial College Healthcare NHS Trust, London, United Kingdom;March of Dimes Prematurity Research Centre, Division of the Institute of Reproductive and Developmental Biology, Department of Metabolism, Digestion, and Reproduction, Imperial College London, London, United Kingdom;St Mary’s Hospital, Imperial College Healthcare NHS Trust, London, United Kingdom;March of Dimes Prematurity Research Centre, Division of the Institute of Reproductive and Developmental Biology, Department of Metabolism, Digestion, and Reproduction, Imperial College London, London, United Kingdom;

关键词: menstrual cup;    swab;    cervicovaginal fluid;    sampling;    microbiota;    metataxonomics;    HIV;   

DOI  :  10.3389/fcimb.2023.1190160

 received in 2023-03-20, accepted in 2023-04-25,  发布年份 2023

来源: Frontiers

【 摘 要 】

BackgroundMenstrual cups (MCs) are increasingly used to collect cervicovaginal secretions to characterise vaginal mucosal immunology, in conjunction with high vaginal swabs (HVS) for metataxonomics, particularly in HIV transmission studies. We hypothesised that both methods of collecting bacterial biomass are equivalent for 16S rRNA gene sequencing.Material and MethodsCervicovaginal fluid (CVF) samples from 16 pregnant women with HIV-1 (PWWH) were included to represent the major vaginal bacterial community state types (CST I-V). Women underwent sampling during the second trimester by liquid amies HVS followed by a MC (Soft disc™) and samples were stored at -80°C. Bacterial cell pellets obtained from swab elution and MC (500 µL, 1 in 10 dilution) were resuspended in 120 µL PBS for DNA extraction. Bacterial 16S rRNA gene sequencing was performed using V1-V2 primers and were analysed using MOTHUR. Paired total DNA, bacterial load, amplicon read counts, diversity matrices and bacterial taxa were compared by sampling method using MicrobiomeAnalyst, SPSS and R.ResultsThe total DNA eluted from one aliquot of diluted CVF from an MC was similar to that of a HVS (993ng and 609ng, p=0.18); the mean bacterial loads were also comparable for both methods (MC: 8.0 log10 16S rRNA gene copies versus HVS: 7.9 log10 16S rRNA gene copies, p=0.27). The mean number of sequence reads generated from MC samples was lower than from HVS (MC: 12730; HVS:14830, p=0.05). The α-diversity metrices were similar for both techniques; MC Species Observed: 41 (range 12-96) versus HVS: 47 (range 16-96), p=0.15; MC Inverse Simpson Index: 1.98 (range 1.0-4.0) versus HVS: 0.48 (range 1.0-4.4), p=0.22). The three most abundant species observed were: Lactobacillus iners, Lactobacillus crispatus and Gardnerella vaginalis. Hierarchical clustering of relative abundance data showed that samples obtained using different techniques in an individual clustered in the same CST group.ConclusionThese data demonstrate that despite sampling slightly different areas of the lower genital tract, there was no difference in bacterial load or composition between methods. Both are suitable for characterisation of vaginal microbiota in PWWH. The MC offers advantages, including a higher volume of sample available for DNA extraction and complimentary assays.

【 授权许可】

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Copyright © 2023 Short, Quinlan, Lee, Preda, Smith, Marchesi, Shattock, Bennett, MacIntyre and Taylor