Significance of Gardnerella vaginalis genotyping in diagnosis of recurrent bacterial vaginosis

2021 ◽  
Vol 1 (30) ◽  
pp. 48-52
Author(s):  
A. A. Krysanova ◽  
A. E. Gushchin ◽  
A. M. Savicheva
Keyword(s):  
Bacterial Vaginosis ◽  
Real Time ◽  
Real Time Pcr ◽  
Test System ◽  
Reproductive Age ◽  
Gardnerella Vaginalis ◽  
First Episode ◽  
Genotype 4 ◽  
Dna Concentration ◽  
Healthy Women

Objective. To assess the importance of identifying different genotypes of Gardnerella vaginalis in the diagnosis of recurrent bacterial vaginosis.Materials and methods. The study involved 299 women of reproductive age. All patients were divided into three groups (healthy women, women with the first episode of bacterial vaginosis, and women with recurrent bacterial vaginosis). DNA of Gardnerella vaginalis in vaginal discharge was detected by real-time PCR. The detection of four genotypes of G. vaginalis was performed using real-time multiplex PCR. To quantify the amplified PCR fragments, quantitative standard samples were constructed. Statistical analysis of the results was carried out using the statistical package NCSS 11 (NCSS, LCC).Results. In 38.2 % of healthy women, any one genotype of G. vaginalis was identified in the vaginal biotope, most often it was genotype 4 (35.2 %), while the concentration of G. vaginalis DNA was low (102–103 geqs/ml). When several genotypes of gardnerella were detected simultaneously in healthy women, the DNA concentration did not exceed 104 geqs/ml. A completely different picture was observed among women with bacterial vaginosis (BV). In the first episode of BV, genotype 4 of G. vaginalis prevailed, both as a single genotype and in combination with 1 or 2, or 3 genotypes. In the recurrent course of BV, only 3–4 genotypes of G. vaginalis were detected at once, and in 78 % of cases it had place is a combination of 1, 2 and 4 genotypes, and the DNA concentration was 107–108 geqs/ml.Conclusion. To diagnose recurrent forms of BV, it is necessary to develop and introduce into practice laboratory diagnostics a test system for detecting different genotypes of G. vaginalis by real-time PCR.

scholarly journals The use of real-time PCR for evaluation of endometrial microbiota

2020 ◽  
pp. 14-20
Author(s):  
E.S. Voroshilina ◽  
D.L. Zornikov ◽  
O.V. Koposova ◽  
D.K. Islamidi ◽  
K.Yu. Ignatova ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Bacterial Load ◽  
Negative Control ◽  
Reproductive Age ◽  
Morphological Pattern ◽  
Chronic Endometritis ◽  
Healthy Women ◽  
Reproductive Age Women ◽  
Lactobacillus Spp

Chronic endometritis (CE) in women of the reproductive age is associated with infertility and recurrent pregnancy loss. The aim of this study was to evaluate the endometrial microbiota by means of real-time PCR in reproductive-age women depending on the morphological pattern of the endometrium. Using the Androflor real-time PCR kit, we analyzed endometrial aspirate collected from 23 patients with chronic endometritis, 30 patients with endometrial hyperplasia, and 19 healthy women. DNA of up to 9 groups of microorganisms was detected in all the analyzed samples in the amounts exceeding negative control. The total bacterial load (TBL) of the detected microorganisms was 10<sup>3</sup>–10<sup>6,4</sup> (median 10<sup>3,8</sup>) GE/ml. Lactobacillus spp. were detected the most often (86.1% of all samples). Opportunistic microorganisms (OM) were identified in 36.1% of all samples, including 22.2% of samples with lactobacilli and 13.9% — without lactobacilli. The variant of microbiota composition with Lactobacillus-dominance (more than 90%. in the TBL) was detected significantly less often in women with chronic endometritis compared to healthy women. Real-time PCR could be used for assessment of endometrial microbiota and allows us to determine its characteristics depending on the morphological pattern.

scholarly journals Quantification of Gardnerella vaginalis, Atopobium vaginae and Lactobacillus spp. in bacterial vaginosis

2021 ◽  
Vol 15 (09) ◽  
pp. 1293-1298
Author(s):  
Nedzib Numanovic ◽  
Snezana Ribis ◽  
Jelena Cukic ◽  
Dane Nenadic ◽  
Aleksandar Zivanovic ◽  
...  
Keyword(s):  
Bacterial Vaginosis ◽  
Real Time ◽  
Real Time Pcr ◽  
Gardnerella Vaginalis ◽  
Pcr Analysis ◽  
Making A Difference ◽  
The Difference ◽  
Lactobacillus Spp ◽  
Atopobium Vaginae

Introduction: The aim of the study was to investigate prevalence of bacteria most frequently associated with bacterial vaginosis using Amsel’s criteria as well as to quantify these bacteria by real-time PCR and to explore the difference in their quantity between healthy and bacterial vaginosis samples. Methodology: For classification of vaginal discharge samples Amsel’s criteria have been used. To detect and quantify Gardnerella vaginalis Atopobium vaginae, Lactobacillus spp. and total vaginal microbiome, real-time PCR has been applied. Results: According to results of our study Amsel’s criteria matched well with real-time PCR diversification of healthy women and women with BV. Nevertheless, real-time PCR has been more sensitive in diagnosis of bacterial vaginosis. DNA quantification of bacteria demonstrated that mutual abundance of G.vaginalis and A. vaginae was good bacterial vaginosis marker . On the contrary, Lactobacillus spp. was present in high amount in both healthy and bacterial vaginosis samples, but ratio of investigated bacteria was different between them. In fact, G. vaginalis and A. vaginae comprised only 0.1% of total microbiome in healthy, whereas Lactobacillus spp. took 99.3% of it. Nonetheless, in bacterial vaginosis, G. vaginalis and A. vaginae made up 34.4% of total microbiome, while Lactobacillus spp. was 21.6%. Conclusions: According to the results of our study real-time PCR analysis was more sensitive in diagnosis of bacterial vaginosis than Amsel’s method, as well as it represented fine tool in making a difference between microbial entities in healthy and bacterial vaginosis samples.

scholarly journals Dynamics of Vaginal Bacterial Communities in Women Developing Bacterial Vaginosis, Candidiasis, or No Infection, Analyzed by PCR-Denaturing Gradient Gel Electrophoresis and Real-Time PCR

2007 ◽  
Vol 73 (18) ◽  
pp. 5731-5741 ◽  
Author(s):  
Beatrice Vitali ◽  
Ciro Pugliese ◽  
Elena Biagi ◽  
Marco Candela ◽  
Silvia Turroni ◽  
...  
Keyword(s):  
Bacterial Vaginosis ◽  
Real Time ◽  
Relative Abundance ◽  
Real Time Pcr ◽  
Bacterial Communities ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Pcr Analysis ◽  
Healthy Women ◽  
Gradient Gel Electrophoresis

ABSTRACT The microbial flora of the vagina plays a major role in preventing genital infections, including bacterial vaginosis (BV) and candidiasis (CA). An integrated approach based on PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and real-time PCR was used to study the structure and dynamics of bacterial communities in vaginal fluids of healthy women and patients developing BV and CA. Universal eubacterial primers and Lactobacillus genus-specific primers, both targeted at 16S rRNA genes, were used in DGGE and real-time PCR analysis, respectively. The DGGE profiles revealed that the vaginal flora was dominated by Lactobacillus species under healthy conditions, whereas several potentially pathogenic bacteria were present in the flora of women with BV. Lactobacilli were the predominant bacterial population in the vagina for patients affected by CA, but changes in the composition of Lactobacillus species were observed. Real-time PCR analysis allowed the quantitative estimation of variations in lactobacilli associated with BV and CA diseases. A statistically significant decrease in the relative abundance of lactobacilli was found in vaginal fluids of patients with BV compared to the relative abundance of lactobacilli in the vaginal fluids of healthy women and patients with CA.

scholarly journals Detection of Human Polyomaviruses in Urine from Bone Marrow Transplant Patients: Comparison of Electron Microscopy with PCR

2004 ◽  
Vol 50 (2) ◽  
pp. 306-312 ◽  
Author(s):  
Stefan S Biel ◽  
Andreas Nitsche ◽  
Andreas Kurth ◽  
Wolfgang Siegert ◽  
Muhsin Özel ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Bone Marrow ◽  
Real Time ◽  
Bone Marrow Transplant ◽  
Real Time Pcr ◽  
Marrow Transplant ◽  
Test System ◽  
Urine Samples ◽  
Clinical Samples ◽  
Transplant Patients

Abstract Background: We studied electron microscopy (EM) as an appropriate test system for the detection of polyomavirus in urine samples from bone marrow transplant patients. Methods: We evaluated direct EM, ultracentrifugation (UC) before EM, and solid-phase immuno-EM (SPIEM). The diagnostic accuracy of EM was studied by comparison with a real-time PCR assay on 531 clinical samples. Results: The detection rate of EM was increased by UC and SPIEM. On 531 clinical urine samples, the diagnostic sensitivity of EM was 47% (70 of 149) with a specificity of 100%. We observed a linear relationship between viral genome concentration and the proportion of urine samples positive by EM, with a 50% probability for a positive EM result for urine samples with a polyomavirus concentration of 106 genome-equivalents (GE)/mL; the probability of a positive EM result was 0% for urine samples with &lt;103 GE/mL and 100% for urine samples containing 109 GE/mL. Conclusions: UC/EM is rapid and highly specific for polyomavirus in urine. Unlike real-time PCR, EM has low sensitivity and cannot quantify the viral load.

scholarly journals Determination of EGFR gene somatic mutations in tissues and plasma of patients with advanced non-small cell lung cancer

2016 ◽  
Vol 62 (6) ◽  
pp. 638-644
Author(s):  
O.I. Brovkina ◽  
M.G. Gordiev ◽  
A.N. Toropovskiy ◽  
D.S. Khodyrev ◽  
R.F. Enikeev ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Small Cell Lung Cancer ◽  
Real Time ◽  
Cell Lung Cancer ◽  
Real Time Pcr ◽  
Test System ◽  
Small Cell ◽  
Egfr Mutations ◽  
Small Cell Lung ◽  
Egfr Gene

The presence of activating mutations in the EGFR gene influences cell proliferation, angiogenesis, and increases metastatic ability; it has a significant impact on the choice of medical therapy of non-small cell lung cancer (NSCLC). The use of targeted therapy with tyrosine kinase inhibitors requires performance of appropriate genetic tests. The aim of this study was to design a real-time PCR-based diagnostic kit for fast and cheap of EGFR mutations testing in paraffin blocks and plasma, and kit validation using samples from patients with NSCLC, and also comparative estimation of diagnostic features of real-time PCR with wild type blocking and digital PCR for mutation testing in blood plasma. The study included 156 patients with various types of adenocarcinoma differentiation. It was designed a simple and efficient real-time PCR-based method of detecting L858R activating mutation and del19 deletion in the EGFR gene for DNA isolated from paraffin blocks. Kit for EGFR mutations was validated using 411 samples of paraffin blocks. The proposed system showed high efficiency for DNA testing from paraffin blocks: a concordance with results of testing with therascreen® EGFR RGQ PCR Kit (`Qiagen`, Germany) was 100%. It has been shown the possibility of using this test system for the detection of mutations in plasma

scholarly journals Age and sex trends of Gardnerella vaginalis infection in patients with sexually transmitted infections in Korea

2021 ◽  
Author(s):  
Young Sam Yuk ◽  
Jae Eun Choi ◽  
Jae Kyung Kim
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Clinical Laboratory ◽  
Gardnerella Vaginalis ◽  
High Tendency ◽  
Sexually Transmitted ◽  
Female Patients ◽  
Average Patient ◽  
Causative Agents ◽  
Age And Sex

Background and Objectives: Gardnerella vaginalis and Candida albicans are the most common causative agents of bac- terial vaginosis, and infections with these pathogens lead to inflammation, endometritis, and pruritus. The aim of this retro- spective study was to determine the trends of G. vaginalis infections based on real-time PCR data according to age and sex in patients with sexually transmitted diseases. Materials and Methods: A total of 59,381 specimens isolated at a clinical laboratory from September 2018 to December 2020 were subjected to real-time PCR for the detection of G. vaginalis DNA. Sample types included catheter, pus, tissue, swab, and urine samples. Results: Among 59,381 samples, 20,718 (34.8%) were positive for G. vaginalis. Of the positive samples, 13,186 (63.7%) were from male patients and 7,532 (36.3%) were from female patients. Average patient age was 39.1 years (the average age of male and female patients was 38.34 and 40.43 years, respectively). Female patients younger than 19 years exhibited the highest incidence of G. vaginalis, at 71.57%, followed by 68.46% incidence in those aged 20-29 years; the lowest incidence was in women aged 40-49 years. Further, among specimen types, the highest number of G. vaginalis-positive specimens was obtained by the swab sampling method. Conclusion: From 2018 to 2020 in Korea, the number of tests conducted for bacterial vaginosis has increased, while the incidence of G. vaginalis infections appears to have decreased. the finding that female adolescents have a high tendency to carry the pathogen is important. and for effective surveillance of BV, sampling by cotton swabs and detection by multiplex PCR might be a good approach.

scholarly journals Prediction of sofosbuvir response using interleukin-6 serum level and single nucleotide polymorphism of interferon lambda- 4

2020 ◽  
Vol 14 (01) ◽  
pp. 80-88
Author(s):  
Amal E Saafan ◽  
Ashraf Abobaker ◽  
Mohamed S Abbas ◽  
Ahmed El-Gendy
Keyword(s):  
Serum Level ◽  
Real Time ◽  
Real Time Pcr ◽  
Dual Therapy ◽  
Snp Genotyping ◽  
Nucleotide Polymorphism ◽  
Time Intervals ◽  
Genotype 4 ◽  
Single Nucleotide ◽  
Taqman Probes

Introduction: In Egypt, 15% of the populations are suffering from chronic hepatitis C especially genotype 4. Sofosbuvir was approved by FDA in December 2013 for treatment of HCV genotypes 2 and 3 in combination with Ribavirin, and for genotypes 1 and 4 in combination with Peg-IFN. Recently, polymorphism of different genes and plasma levels of IL-6 were utilized for better prediction of HCV clearance. This study aimed at early prediction of the efficacy of HCV treatment with Sofosbuvir (Sovaldi) and comparing the antiviral efficacy of dual and triple Sovaldi combination therapy. Methodology: Blood samples were collected from 100 HCV positive patients and detected by real time PCR at three time intervals. SNP genotyping of INFL-4 gene was estimated by using real-time PCR with predesigned primers and Taqman probes. IL-6 serum level was estimated before, during and after the end of the treatment using ELISA assay based on human IL-6 KIT. Results: SNP genotyping of INFL-4 gene showed that 13.1% of patients carried ∆G/∆G, 30.4% patients had TT/TT and 56.5% patients possessed heterozygote allele ∆G/TT. Clinical data displayed that 13 patients were got relapsed at SVR 12. Serum level of IL-6 was noticed higher in HCV patients than healthy ones. Noteworthy, it was increased during treatment then decreased to a minimal level than begining of treatment. Conclusion: SNP in INFL-4 gene has displayed no effect in response to Sofosbuvir. Dual therapy had the same effect like triple therapy, so interferon could be withdrawn from the treatment regimen.

scholarly journals Correction for Hilbert et al., Development and Validation of a Highly Accurate Quantitative Real-Time PCR Assay for Diagnosis of Bacterial Vaginosis

2016 ◽  
Vol 54 (7) ◽  
pp. 1930-1930 ◽  
Author(s):  
David W. Hilbert ◽  
William L. Smith ◽  
Sean G. Chadwick ◽  
Geoffrey Toner ◽  
Eli Mordechai ◽  
...  
Keyword(s):  
Bacterial Vaginosis ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Real Time Pcr ◽  
Pcr Assay ◽  
Development And Validation

USE OF FEMOFLOR-16 TEST TO ASSESS GENITAL BIOCENOSIS IN WOMEN WITH INFLAMMATORY AND PROLIFERATIVE DISEASES OF CERVIX

2016 ◽  
Vol 1 (61) ◽  
pp. 90-95
Author(s):  
Иевлева ◽  
Nadezhda Ievleva ◽  
Пермина ◽  
Natalya Permina ◽  
Ивахнишина ◽  
...  
Keyword(s):  
Genital Tract ◽  
Inflammatory Diseases ◽  
Female Genital Tract ◽  
Reproductive Age ◽  
Gardnerella Vaginalis ◽  
Control Group ◽  
Healthy Women ◽  
Opportunistic Bacteria ◽  
Obligate Anaerobic ◽  
Atopobium Vaginae

Qualitative and quantitative assessment of microbes making the microbiocenosis of genital tract in women with inflammatory and proliferative diseases of cervix using Femoflor-16 test was the aim of the research. Scrapings of cervix and vaginal fornix in 100 women of reproductive age with cervicitis, vaginitis and in 31 women with cervical pseudoerosion (ectropion) were studied. The control group consisted of 35 relatively healthy women preparing for pregnancy. Cervical and vaginal dysbiosis was found in women with inflammatory diseases of cervix in 37.0% of cases, in women with pseudoerosion in 32.2% of cases. These are 3.3 (р&#60;0.005) and 2.8 (р&#60;0.02) times as much as in the group of healthy women preparing for pregnancy (11.1%). Dysbiosis structure was represented primarily by obligate anaerobic agents such as Gardnerella vaginalis, Atopobium vaginae, Eubacterium spp. in association with other opportunistic bacteria that are clinically most significant microorganisms colonizing female genital tract. Mycoplasma and yeast-like fungi of Candida species were found primarily with anaerobes. Aerobic and mixed dysbiosis were only found in 7% of cases in women with inflammatory diseases. Femoflor-16 test is a readily available, fast, efficient, up-to-date method enabling one to begin with early adequate antibacterial therapy and monitor it.

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