Vaginal Microbiota Composition Correlates Between Pap Smear Microscopy and Next Generation Sequencing and Associates to Socioeconomic Status

ABSTRACTDeep characterization, even by next-generation sequencing, of the vaginal microbiota in healthy women or posttreatment bacterial vaginosis patients is limited by the dominance of lactobacilli. To improve detection, we offer two approaches: quantitative PCR (qPCR) using phylogenetic branch-inclusive primers and sequencing of broad-spectrum amplicons generated with oligomers that block amplification of lactobacilli.

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Determination of community state types of vaginal microbiota in pregnant Korean women by using next-generation sequencing

European Journal of Obstetrics & Gynecology and Reproductive Biology ◽

10.1016/j.ejogrb.2018.08.430 ◽

2019 ◽

Vol 234 ◽

pp. e123

Author(s):

Kwan Young Oh ◽

Eun Jung Shim ◽

In Young Kim ◽

Chan Hee Jin

Keyword(s):

Next Generation Sequencing ◽

Vaginal Microbiota ◽

Korean Women ◽

Next Generation ◽

Generation Sequencing

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The association between gut microbiota composition and BMI in Chinese male college students, as analysed by next-generation sequencing

British Journal Of Nutrition ◽

10.1017/s0007114519001909 ◽

2019 ◽

Vol 122 (9) ◽

pp. 986-995 ◽

Cited By ~ 1

Author(s):

Yanrong Lv ◽

Xiangxiang Qin ◽

Huaijie Jia ◽

Sirui Chen ◽

Weiwei Sun ◽

...

Keyword(s):

College Students ◽

Next Generation Sequencing ◽

Gut Microbiota ◽

Microbiota Composition ◽

Next Generation ◽

Male College ◽

Chinese Male ◽

Generation Sequencing ◽

Gut Microbiota Composition ◽

Male College Students

AbstractAltered gut microbial ecology contributes to the development of metabolic diseases including obesity. However, studies based on different populations have generated conflicting results due to diet, environment, methodologies, etc. The aim of our study was to explore the association between gut microbiota and BMI in Chinese college students. The 16S next-generation sequencing (NGS) was used to test the gut microbiota of nine lean, nine overweight/obesity and ten normal-weight male college students. The differences in gut microbiota distribution among three groups were compared, and the relationship between the richness, diversity, composition of gut microbiota and BMI were analysed. The predominant phyla Bacteroidetes and Firmicutes were further confirmed by real-time PCR. Metagenomic biomarker discovery was conducted by linear discriminant analysis (LDA) effect size (LEfSe). NGS revealed that gut microbiota composition was different among three groups, but there was no difference in the abundance ratio of Firmicutes:Bacteroidetes. Several bacterial taxa were in linear relationship with BMI (positive relationship: uncultured bacterium (Bacteroides genus); negative relationship: Porphyromonadaceae, Acidaminococcaceae, Rikenellaceae, Desulfovibrionaceae, Blautia, Anaerotruncus, Parabacteroides, Alistipes). Moreover, gut microbiota diversity decreased with the increase in BMI. And LEfSe analysis indicated that Blautia, Anaerotruncus and its uncultured species were significantly enriched in the lean group (LDA score ≥ 3), Parasuterella and its uncultured species were significantly enriched in the overweight/obese groups (LDA score ≥ 3). In general, gut microbiota composition and microbial diversity were associated with BMI in Chinese male college students. Our results might enrich the understanding between gut microbiota and obesity.

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Comparison of different hypervariable regions of 16S rRNA for taxonomic profiling of vaginal microbiota using next-generation sequencing

Archives of Microbiology ◽

10.1007/s00203-020-02114-4 ◽

2020 ◽

Author(s):

Auttawit Sirichoat ◽

Nipaporn Sankuntaw ◽

Chulapan Engchanil ◽

Pranom Buppasiri ◽

Kiatichai Faksri ◽

...

Keyword(s):

Next Generation Sequencing ◽

16S Rrna ◽

Vaginal Microbiota ◽

Next Generation ◽

Taxonomic Profiling ◽

Hypervariable Regions ◽

Generation Sequencing

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O-130 The influence of vaginal microbiota on frozen blastocyst implantation after transfer: a prospective study through next-generation 16S rRNA sequencing

Human Reproduction ◽

10.1093/humrep/deab126.055 ◽

2021 ◽

Vol 36 (Supplement_1) ◽

Author(s):

H Asakura ◽

Y Nakahara ◽

Y Nagai ◽

Y Sakuraba

Keyword(s):

Next Generation Sequencing ◽

16S Rrna ◽

Prospective Study ◽

Embryo Implantation ◽

Vaginal Microbiota ◽

Vaginal Fluid ◽

Next Generation ◽

The Difference ◽

A Prospective Study ◽

Generation Sequencing

Abstract Study question A prospective study to investigate the relationship between the composition of vaginal microbiota through next-generation sequencing and the efficacy of single frozen blastocyst transfer in the same cycle. Summary answer Dominant presence of lactobacillus and other lactate producing microbes in the upper vagina was highly correlated with implantation of transferred blastocyst in this pilot study. What is known already Next-generation sequencing of 16S rRNA detected microbes in the uterine cavity and recent studies indicated that dominant presence of Lactobacillus correlated highly with successful implantation of the transferred embryos. Aberrant vaginal microbiota has been known to cause poor obstetrical outcomes, however little is known for its effect on embryo implantation in assisted reproduction. Study design, size, duration A prospective study with 25 female subjects transferring a frozen blastocyst using autologous oocyte, over 14 months period in 2019-2020. Participants/materials, setting, methods 25 female patients without tubal and uterine pathology and no history of multiple miscarriages and implantation failures were recruited with consent at a private ART clinic. Transdermal estrogen was used to prepare endometrium. Upper vaginal fluid was obtained in follicular phase of the the same cycle and analyzed through next-generation sequencing, but the result was reported after pregnancy confirmation. Single frozen blastocyst transfer and standard luteal phase support were performed. Institutional IRB approved the protocol. Main results and the role of chance The mean age was 36.2 y.o.(range 29-41 y.o.), and 14 gestational sacs (implantation rate 56%), and 3 miscarriage (21.4%) were observed. Next-generation sequencing for 16S rRNA revealed average 69.2% presence of Lactobacillus (0-100%) and average 78.0% (0.2-100%) lactate producing microbes (LPM: Lactobacillus, Bifidobacterium, Streptococcus, and Enterococcus) in the vaginal fluid. Using 90% as cut-off, implantation rates were 11/15 (73.3%) and 3/10 (30%) with Lactobacillus dominant and non-dominant, 12/16 (75%) and 2/9 (22.2%) with LPM dominant and non-dominant microbiota, respectively. The difference in each group were not statistically significant. The relative risks for pregnancy were 2.63 (95%CI 1.03-6.67, P=0.04) for Lactobacillus and 3.11 (95%CI 1.24-7.79, P=0.02) for LPM. As for ROC analysis for embryo implantation and dominant microbes, AUC and associated criterion were 0.62 and 90.7% (sensitivity 78.6%, specificity 72.7%) for Lactobacillus, 0.69 and 96.6% (sensitivity 85.7%, specificity 72.7%) for LPM, respectively. The difference of AUC was not significant (P=0.24). Limitations, reasons for caution Despite prospective nature of the study, small sample size limited the analytical power of the study. Aneuploidy screening was not performed to remove confounding factor. Wider implications of the findings Our pilot study revealed possible relationship between vaginal microbiota and embryo implantation. Dominance of Lactobacillus or other lactate producing microbes may be advantageous for successful ART. Sampling vaginal fluid for microbe analysis is less invasive than endometrial sampling and can obtain more abundant RNA with possible higher accuracy of analysis. Trial registration number not applicable

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The vaginal microbiome of sub-Saharan African women: revealing important gaps in the era of next-generation sequencing

PeerJ ◽

10.7717/peerj.9684 ◽

2020 ◽

Vol 8 ◽

pp. e9684

Author(s):

Nkechi Martina Odogwu ◽

Oladapo O. Olayemi ◽

Akinyinka O. Omigbodun

Keyword(s):

Next Generation Sequencing ◽

Large Scale ◽

Marker Gene ◽

Vaginal Microbiota ◽

African Population ◽

African Women ◽

Published Data ◽

Vaginal Microbiome ◽

Next Generation ◽

Generation Sequencing

Accurate characterization of the vaginal microbiome remains a fundamental goal of the Human Microbiome project (HMP). For over a decade, this goal has been made possible deploying high-throughput next generation sequencing technologies (NGS), which indeed has revolutionized medical research and enabled large-scale genomic studies. The 16S rRNA marker-gene survey is the most commonly explored approach for vaginal microbial community studies. With this approach, prior studies have elucidated substantial variations in the vaginal microbiome of women from different ethnicities. This review provides a comprehensive account of studies that have deployed this approach to describe the vaginal microbiota of African women in health and disease. On the basis of published data, the few studies reported from the African population are mainly in non-pregnant post pubertal women and calls for more detailed studies in pregnant and postnatal cohorts. We provide insight on the use of more sophisticated cutting-edge technologies in characterizing the vaginal microbiome. These technologies offer high-resolution detection of vaginal microbiome variations and community functional capabilities, which can shed light into several discrepancies observed in the vaginal microbiota of African women in an African population versus women of African descent in the diaspora.

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Next-Generation Sequencing as a Tool to Detect Vaginal Microbiota Disturbances during Pregnancy

Microorganisms ◽

10.3390/microorganisms8111813 ◽

2020 ◽

Vol 8 (11) ◽

pp. 1813 ◽

Cited By ~ 1

Author(s):

Agnieszka Sroka-Oleksiak ◽

Tomasz Gosiewski ◽

Wojciech Pabian ◽

Artur Gurgul ◽

Przemysław Kapusta ◽

...

Keyword(s):

Next Generation Sequencing ◽

Streptococcus Agalactiae ◽

Vaginal Microbiota ◽

Quantitative Composition ◽

Next Generation ◽

Lactobacillus Jensenii ◽

Sequencing Method ◽

Genus Lactobacillus ◽

Caucasian Women ◽

Generation Sequencing

The physiological microbiota of the vagina is responsible for providing a protective barrier, but Some factors can disturb the balance in its composition. At that time, the amounts of the genus Lactobacillus decrease, which may lead to the development of infection and severe complications during pregnancy. The aim of the study was the analysis of the bacterial composition of the vagina in 32 Caucasian women at each trimester of pregnancy using the next-generation sequencing method and primers targeting V3-V4 regions. In the studied group, the dominant species were Lactobacillus iners, Lactobacillus gasseri, and Lactobacillusplantarum. Statistically significant differences in the quantitative composition between trimesters were observed in relation to Lactobacillus jensenii,Streptococcus agalactiae, Lactobacillus iners, Gardnerella spp. Out of the 32 patients, 20 demonstrated fluctuations within the genus Lactobacillus, and 9 of them, at different stages of pregnancy, exhibited the presence of potentially pathogenic microbiota, among others: Streptococcus agalactiae, Gardnerella spp., Atopobium vaginae, and Enterococcus faecalis. The composition of the vaginal microbiota during pregnancy was subject to partial changes over trimesters. Although in one-third of the studied patients, both the qualitative and quantitative composition of microbiota was relatively constant, in the remaining patients, physiological and potentially pathogenic fluctuations were distinguished.

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