Molecular Detection and Genotyping of Gardnerella Vaginalis, 16S rRNA Gene from Bacterial Vaginosis Miscarriage Women in AL-Hillah City

2020 ◽  
Vol 24 (5) ◽  
pp. 1536-1544
Author(s):  
Asmaa K. Gatea
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Bacterial Vaginosis ◽  
Molecular Detection ◽  
Gardnerella Vaginalis ◽  
Rrna Gene

scholarly journals 2576. The Microbiome of Recurrent Bacterial Vaginosis Compared with Asymptomatic Controls

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S895-S895
Author(s):  
Elizabeth O Shay ◽  
Oluwatosin Goje ◽  
Roshan Padmanabhan ◽  
Charis Eng
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Bacterial Vaginosis ◽  
Alpha Diversity ◽  
Dna Probe ◽  
Gardnerella Vaginalis ◽  
Rrna Gene ◽  
Vaginal Microbiome ◽  
Β Diversity ◽  
Probe Test

Abstract Background Bacterial vaginosis (BV) affects nearly 1 in 3 women in the United States and is poorly understood. The study of the vaginal microbiome, using 16S rRNA-gene amplicon sequencing, has increased our knowledge of BV. We aimed to characterize the vaginal microbiome of women with recurrent BV firstly in comparison to controls, and secondly in comparison to a sub-population of our asymptomatic controls, positive for Gardnerella vaginalis via a vaginal pathogens DNA direct probe test (DNA probe). Methods Women aged 18–40 years, with recurrent BV, and asymptomatic controls were prospectively enrolled. Vaginal samples were collected from each participant. DNA was extracted, amplified using primers targeting the V3-V4 variable region of the 16S rRNA-gene, and then sequenced and processed through a hybrid Qiime MICCA bioinformatics pipeline. We also tested for G. vaginalis using the DNA probe. Results Seventeen recurrent BV patients and 46 controls were enrolled. Β diversity (P = 0.045), but not alpha diversity (P = 0.076) differed between groups. The genera Gardnerella and Prevotella were relatively more abundant, while Lactobacillus was relatively less abundant in recurrent BV vs. control groups. Of the patients for whom results of the DNA probe for Gardnerella vaginalis were available, 11 (69%) recurrent BV patients and 14 (35%) controls were positive. Control patients, negative by the DNA probe test, showed decreased alpha diversity (P = 0.0001) and significantly different β diversity (P = 0.001) compared with recurrent BV patients. Neither alpha (P = 0.31) nor β (P = 0.096) diversity differed between recurrent BV patients and controls that were G. vaginalis positive. Conclusion The microbiome of recurrent BV patients is distinct from that of asymptomatic controls; recurrent BV patients exhibit different β diversity, less Lactobacillus and more Gardnerella and Prevotella. Asymptomatic Gardnerella vaginalis-colonized controls demonstrate similar microbiome profiles to those of recurrent BV patients. These findings suggest that individual factors may influence whether or not a patient with a BV microbiomic profile experiences symptoms. Further investigation into these mechanisms could yield insights into the treatment of recurrent BV. Disclosures All authors: No reported disclosures.

scholarly journals Suitability of a Unique 16S rRNA Gene PCR Product as an Indicator of Gardnerella vaginalis

BioTechniques ◽  
2000 ◽  
Vol 28 (2) ◽  
pp. 222-226 ◽  
Author(s):  
Kamalendu Nath ◽  
Joseph W. Sarosy ◽  
Spyros P. Stylianou
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Gardnerella Vaginalis ◽  
Rrna Gene ◽  
Pcr Product

scholarly journals Molecular detection of Anaplasmataceae agents in Dasyprocta azarae in northeastern Brazil

2018 ◽  
Vol 27 (1) ◽  
pp. 98-104 ◽  
Author(s):  
Maria do Socorro Costa de Oliveira Braga ◽  
José Gomes Pereira ◽  
Simone de Jesus Fernandes ◽  
Ingrid Carolinne Lopes Marques ◽  
Renata Passos de Jesus ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Molecular Detection ◽  
Evolutionary Model ◽  
Northeastern Brazil ◽  
Likelihood Method ◽  
Rrna Gene ◽  
Nested Pcr Assays ◽  
Tick Vectors ◽  
The 16S Rrna Gene

Abstract Recently, the importance of wild-living rodents for maintenance of pathogens of the family Anaplasmataceae in the environment was investigated. These mammals play a role as reservoirs for these pathogens and act as hosts for the immature stages of tick vectors. The aim of the present study was to investigate the prevalence of Ehrlichia sp. and Anaplasma sp. in 24 specimens of Azara’s agouti (Dasyprocta azarae) that had been trapped in the Itapiracó Environmental Reserve, in São Luís, Maranhão, northeastern Brazil, using molecular methods. Four animals (16.7%) were positive for Ehrlichia spp. in nested PCR assays based on the 16S rRNA gene. In a phylogenetic analysis based on the 16S rRNA gene, using the maximum likelihood method and the GTRGAMMA+I evolutionary model, Ehrlichia sp. genotypes detected in Azara’s agoutis were found to be closely related to E. canis and to genotypes relating to E. canis that had previously been detected in free-living animals in Brazil. The present work showed the first molecular detection of Ehrlichia sp. in Azara’s agoutis in Brazil.

scholarly journals MOLECULAR IDENTIFICATION AND ANTIBIOGRAM PROFILES OF ESCHERICHIA COLI ISOLATED FROM APPARENTLY HEALTHY AND DIARRHEIC GOATS

2017 ◽  
Vol 14 (2) ◽  
pp. 203-208 ◽  
Author(s):  
F. Begum ◽  
M. M. Islam ◽  
M. Sohidullah ◽  
S. M. L. Kabir ◽  
M. Islam ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Teaching Hospital ◽  
Molecular Detection ◽  
Biochemical Characterization ◽  
Rrna Gene ◽  
E Coli ◽  
Goat Farm ◽  
Apparently Healthy

The present study was designed for the cultural, biochemical characterization and molecular detection of E. coli from apparently healthy and diarrheic goats in and around BAU campus including their antibiogram study. A total of 50 fecal samples were collected among which 13 originated from diarrheic goat and 37 from apparently healthy goats. Out of 50 samples, 35 were found positive for E. coli i.e., overall 70% occurrence. Occurrences of E. coli from diarrheic and apparently healthy goats were 92% and 62% respectively. Occurrences were 60%, 80% and 70% in case of BAU Goat Farm, Veterinary Teaching Hospital and Boyra respectively. On age basis 93%, 54%, 66% and 54% samples originated from 6 months, 7-12 months, 13-18 months and 19 months aged goats were found positive respectively. Occurrences of E. coli on the basis of sex were 78% for male and 62% for female. In case of breed, the occurrences were 69% in Black Bengal and 100% in for Jamunapari. Molecular detection was done by PCR and 13 out of 20 isolates tested gave the bands at the 585 bp specific for E. coli 16S rRNA gene. All the isolates (100%) were found sensitive to ciprofloxacin and norfloxacin; 100% and 35% were intermediately resistant to tetracycline and gentamicin respectively and 25% isolates were resistant to streptomycin. Ciprofloxacin and norfloxacin were found to be the best choice of antibiotics for the treatment of colibacillosis in goats in the study area.

scholarly journals Complementing 16S rRNA gene amplicon sequencing with estimates of total bacterial load to infer absolute species concentrations in the vaginal microbiome

2019 ◽  
Author(s):  
Florencia Tettamanti Boshier ◽  
Sujatha Srinivasan ◽  
Anthony Lopez ◽  
Noah G. Hoffman ◽  
Sean Proll ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Bacterial Vaginosis ◽  
Relative Abundance ◽  
Bacterial Species ◽  
Bacterial Load ◽  
Amplicon Sequencing ◽  
Individual Species ◽  
Rrna Gene ◽  
Associated Bacteria

Whereas 16S rRNA gene amplicon sequencing quantifies relative abundances of bacterial taxa, variation in total bacterial load between samples restricts its ability to reflect absolute concentration of individual species. Quantitative PCR (qPCR) can quantify individual species, but it is not practical to develop a suite of qPCR assays for every bacterium present in a diverse sample. We analyzed 1320 samples from 20 women with a history of frequent bacterial vaginosis, who self-collected vaginal swabs daily over 60 days. We inferred bacterial concentrations by taking the product of species relative abundance (assessed by 16S rRNA gene amplicon sequencing) and total bacterial load (measured by broad-range 16S rRNA gene qPCR). Log10-converted inferred concentrations correlated with targeted qPCR (r = 0. 935, p<2.2e-16) for seven key bacterial species. The mean inferred concentration error varied across bacteria, with rarer bacterial vaginosis-associated bacteria associated with larger errors. 92% of errors >0.5 log10 occurred when relative abundance was <10%. Many errors occurred during early bacterial expansion or late contraction. When relative abundance of a species is >10%, inferred concentrations are reliable proxies for targeted qPCR. However, targeted qPCR is required to capture bacteria at low relative abundance, particularly with BV-associated bacteria during the early onset of bacterial vaginosis.

scholarly journals Molecular Detection of Campylobacter spp. in California Gull (Larus californicus) Excreta

2011 ◽  
Vol 77 (14) ◽  
pp. 5034-5039 ◽  
Author(s):  
Jingrang Lu ◽  
Hodon Ryu ◽  
Jorge W. Santo Domingo ◽  
John F. Griffith ◽  
Nicholas Ashbolt
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Gene Sequence ◽  
Molecular Detection ◽  
Rrna Gene ◽  
Fecal Indicators ◽  
Sequencing Data ◽  
Content Type ◽  
Human Illness ◽  
Campylobacter Spp

ABSTRACTWe examined the prevalence, quantity, and diversity ofCampylobacterspecies in the excreta of 159 California gull (Larus californicus) samples using culture-, PCR-, and quantitative PCR (qPCR)-based detection assays.Campylobacterprevalence and abundance were relatively high in the gull excreta examined; however,C. jejuniandC. lariwere detected in fewer than 2% of the isolates and DNA extracts from the fecal samples that tested positive. Moreover, molecular and sequencing data indicated that mostL. californicuscampylobacters were novel (<97% 16S rRNA gene sequence identity to knownCampylobacterspecies) and not closely related to species commonly associated with human illness.Campylobacterestimates were positively related with those of fecal indicators, including a gull fecal marker based on theCatellicoccus marimammalium16S rRNA gene.

scholarly journals Prevalence and Abundance of Uncultivated Megasphaera-Like Bacteria in the Human Vaginal Environment

2008 ◽  
Vol 74 (5) ◽  
pp. 1656-1659 ◽  
Author(s):  
Marcela Zozaya-Hinchliffe ◽  
David H. Martin ◽  
Michael J. Ferris
Keyword(s):  
Phylogenetic Analysis ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Bacterial Vaginosis ◽  
Quantitative Pcr ◽  
Rrna Gene ◽  
Gene Sequences ◽  
16S Rrna Gene Sequences ◽  
Environmental Distribution ◽  
Independent Analysis

ABSTRACT Cultivation-independent analysis of 16S rRNA gene sequences in vaginal samples revealed two previously unrecognized, uncultivated Megasphaera-like phylotypes. Phylogenetic analysis and environmental distribution suggest that these Megasphaera types may be unique to the vaginal environment. Quantitative PCR suggests that both phylotypes are present in higher concentrations in women with bacterial vaginosis.

scholarly journals ANTIBIOGRAM AND MOLECULAR CHARACTERIZATION OF VIBRIO CHOLERAE ISOLATED FROM MARINE FISH

2017 ◽  
Vol 10 (6) ◽  
pp. 146
Author(s):  
Rwarinda U Angelo ◽  
Samiraj Ramesh Ramesh
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Vibrio Cholerae ◽  
Marine Fish ◽  
Microscopic Observation ◽  
Molecular Detection ◽  
Pcr Amplification ◽  
Diffusion Method ◽  
Rrna Gene ◽  
Biochemical Tests

Objective: Molecular identification and antibiotic susceptibility evaluation of Vibrio cholerae from marine fish available in local fish market Thanjavur, Tamil nadu, India.Methods: inoculation was done by using nutrient agar as general media and TCBS agar as selective media and confirmed as V.cholerae by Gram stain (Microscopic Observation), Growth characteristics of different media, Biochemical tests like Methyl Red Test, Nitrate Reduction Tests, Indole Test etc. Sensitivity (drug sensitivity) was done in Muller Hinton Agar (MH Agar) using disc diffusion method ten different antibiotics were used to evaluate the antibiogram profile, molecular detection was done by targeting 16S rRNA gene by using a universal primer.Results:  V.cholerae is present in marine fish samples, as showed by culture method and microscopic observation as well biochemical tests. PCR amplification of 16S rRNA gene showed the amplification of targeted gene and antibiogram profile showed that isolates are more sensitive to Ampicillin in comparison with others antibiotics used in this study. Ampicillin can be used for V.cholerae infection by the physicians and amoxicillin must be avoided which is resistant.Conclusion: Molecular detection is safe and rapid methods for bacteria identification as revealed by PCR amplification of 16S rRNA gene. As the isolates are more sensitive to Ampicillin in comparison with others antibiotics used in this study. Ampicillin can be used for V.cholerae infection by the physicians and amoxicillin and nitrofurantoin must be avoided.      

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