scholarly journals Distribution of polymorphic variants of genes for xenobiotic biotransformation GSTM1, GSTT1 and GSTP1 in populations of native inhabitants and Russians of Eastern Siberia

2020 ◽  
pp. 24-34
Author(s):  
Л.Э. Табиханова ◽  
Л.П. Осипова ◽  
Е.Н. Воронина ◽  
М.Л. Филипенко
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Genetic Research ◽  
Total Sample ◽  
Eastern Siberia ◽  
Multifactorial Diseases ◽  
Genotypic Combination ◽  
Risk Alleles ◽  
Polymorphic Variants ◽  
The Difference

Актуальность. Изучение полиморфизма генов системы биотрансформации ксенобиотиков, ассоциированных с рядом многофакторных заболеваний - важное направление современных медико-генетических исследований. Цель и задачи - выявить этнические особенности распределения полиморфных вариантов генов GSTM1, GSTT1 и GSTP1 среди бурят, телеутов и русских Восточной Сибири. Материалы и методы. Изучены выборки восточных (N=139) и западных (N=284) бурят, метисов западных бурят с русскими (N=47), телеутов (N=115) и русских Восточной Сибири (N=122). Выявление генотипов GSTM1 0/0 и GSTT1 0/0 проводилось методом мультиплексной полимеразной цепной реакции в режиме реального времени, генотипирование GSTP1 проводили в режиме реального времени с использованием TaqMan-зондов. Результаты. Встречаемость генотипа GSTM1 0/0 среди восточных и западных бурят составляет 37,7% и 57,7% соответственно (51,4% в суммарной выборке бурят), среди русских - 42,6%. Статистически значимо меньшая частота показана у телеутов - 17,4%. Частота GSTТ1 0/0 у восточных и западных бурят равна 40,8% и 27,6% соответственно, у русских статистически значимо меньше - 18%, у телеутов - 24,8%. Для метисов показаны промежуточные значения частот GSTM1 0/0 и GSTТ1 0/0. Аллель GSTP1 1405G встречается среди восточных и западных бурят с частотой 27,7% и 19,2% соответственно, у русских - 31,8%, телеутов - 24,8%. Различие русских с западными бурятами статистически значимо. Частота аллеля GSTP1 2285T среди восточных (4,9%), западных (1,8%) бурят и телеутов (2,2%) меньше, чем среди русских (8,3%). Отличие русских от западных бурят и телеутов, является статистически значимым. Выводы. В суммарной выборке бурят показаны повышенные частоты генотипов GSTM1 0/0 и GSTТ1 0/0, ассоциированых, по данным литературы, с некоторыми многофакторными заболеваниями по сравнению с телеутами и русскими. В обеих выборках бурят статистически значимо повышена частота комбинированного генотипа, приводящего к отсутствию активности ферментов. В то же время у телеутов частота индивидов с генотипической комбинацией GSTM1 +GSTТ1 +, ответственной за нормальную ферментативную активность, статистически значимо выше. Частоты аллелей 1405G и 2285T гена GSTP1 среди бурят и телеутов понижены по сравнению с русскими. Метисация способствует изменению частоты аллелей. Статистически значимые различия в частотах вариантов GSTM1, GSTТ1 и GSTP1 внутри бурятского этноса могут свидетельствовать о его генетической неоднородности. Relevance. The study of the gene polymorphism of the system biotransformation of xenobiotics, which are associated with a number of multifactorial diseases, is an important area of modern medical genetic research. The aim and tasks are to reveal ethnic features in the distribution of polymorphic variants of genes GSTM1, GSTT1 and GSTP1 among Buryats, Teleuts and Russians of Eastern Siberia. Materials and methods. The samples of Eastern (N=139) and Western (N=284) Buryats, métis Western Buryats with Russians (N=47), Teleuts (N=115) and Russians of East Siberia (N=122) are studied. Detection of GSTM1 0/0 and GSTT1 0/0 was carried out through multiplex real-time PCR. Genotyping of GSTP1 was performed using TaqMan real-time PCR. Results. The occurrence of GSTM1 0/0 among Eastern and Western Buryats is 37.7% and 57.7% respectively (51.4% for the total sample of Buryats), among Russians - 42.6%. Significantly lower frequency is shown in Teleuts-17.4%. The frequency of GSTT1 0/0 in Eastern and Western Buryats is 40.8% and 27.6% respectively. It is significantly lower in Russians - 18%, in Teleuts - 24.8%. Mestis shows intermediate frequencies of GSTM1 0/0 and GSTТ1 0/0. GSTP1 1405G is found among Eastern and Western Buryats with 27.7% and 19.2% frequency, respectively, in Russians - 31.8%, Teleuts - 24.8%. The difference of Russians with Western Buryats is statistically significant. The frequencies of GSTP1 2285T among Eastern (4.9%), Western (1.8%) Buryats and Teleuts (2.2%) are lower than frequency among Russians (8.3%). The difference between Russians and Western Buryats with Teleuts is statistically significant. Summary. There are increased frequencies of GSTM1 0/0 and GSTT1 0/0 in the total cohort of Buryats in comparison with Teleuts and Russians. According to the literature data, these genotypes are associated with multi-factorial diseases. In both samples of Buryats, there is a statistically significantly increased frequency of the combined genotype resulting in the absence of enzyme activity. At the same time, there is a statistically significantly increased frequency of individuals with a genotypic combination GSTM1 +GSTT1 + responsible for normal enzymatic activity in the sample of Teleuts. There are reduced frequencies of risk-alleles GSTP1 1405G and 2285T among Buryats and Teleuts in comparison with Russians. Metisation changes the frequency of risk alleles. Significant differences in the frequencies of GSTM1, GSTT1 and GSTP1 within the Buryat ethnic group may indicate its genetic heterogeneity.

scholarly journals Detection of soybean by real-time PCR in the samples subjected to deep technological processing

2019 ◽  
Vol 4 (4) ◽  
pp. 23-27
Author(s):  
K. A. Kurbakov ◽  
E. A. Konorov ◽  
V. N. Zhulinkova ◽  
M. Yu. Minaev
Keyword(s):  
Glycine Max ◽  
Real Time ◽  
Real Time Pcr ◽  
High Sensitivity ◽  
Food Product ◽  
Single Copy ◽  
Probe System ◽  
Canned Foods ◽  
Technological Processing ◽  
The Difference

During deep technological processing, DNA of food product components (specifically, in canned foods) is subjected to strong degradation, which makes the PCR-based food components identification more difficult. In this work, a primer-probe system is proposed, which was selected for the multi-copy region of long terminal repeat (LTR) of soybean (Glycine max). We confirmed its high sensitivity and specificity for soybean detection by real-time PCR. Using the selected system, we successfully analyzed the samples of meat-and-plant canned foods and other food products subjected to deep technological processing — tofu, preserved tofu, soy sauces, confectionary products containing soy lecithin. To compare with these samples, real-time PCR was carried out using the primer-probe system selected for the single-copy le1 gene. In terms of sensitivity, the use of the primer-probe system specific to the single-copy region was significantly inferior to the primer-probe system specific to the LTR region. The difference in the rate of degradation of these genomic DNA regions of Glycine max was found.

Loss of Proapoptotic Gene Apaf-1 Expression in Pancreatic Cancer

2015 ◽  
Vol 25 (2) ◽  
pp. 9-12
Author(s):  
Albertas Daukša ◽  
Aldona Jakštaitė ◽  
Augustina Gasianec ◽  
Žilvinas Dambrauskas
Keyword(s):  
Pancreatic Cancer ◽  
Mrna Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Pathological Features ◽  
Cellular Apoptosis ◽  
The Difference ◽  
Proapoptotic Gene

Pancreatic cancer has one of the worst prognoses of allgastrointestinal malignancies. One of the key proteins in themechanism of apoptosis is apoptosis protease-activating factorAPAF-1. The aim of this study was to determine expression ofAPAF-1 mRNA in pancreatic cancer. Real time PCR for APAF-1 mRNA was performed on 29 pancreatic cancer cases. Theassociation of APAF-1 mRNA expression with clinico-pathological features was analysed. APAF-1 mRNA expression was lower in 82.8 % in pancreatic cancer than in healthy tissues. The difference was found statistically significant (p=0.02). No association was found between APAF-1 mRNA expression in cancerous tissues and clinico-pathological features. To conclude, in this study we have demonstrated mRNA decrease of APAF-1, indicating that gene involved in cellular apoptosis might be involved in pancreaticcancer pathogenesis.

Using genes differentially expressed in bulls to classify steers divergently selected for high and low residual feed intake

2012 ◽  
Vol 52 (7) ◽  
pp. 608 ◽  
Author(s):  
Y. Chen ◽  
P. F. Arthur ◽  
R. M. Herd ◽  
K. Quinn ◽  
I. M. Barchia
Keyword(s):  
Real Time ◽  
Feed Intake ◽  
Real Time Pcr ◽  
Calcium Binding ◽  
Correlation Coefficients ◽  
Residual Feed Intake ◽  
Differentially Expressed ◽  
Quantitative Real Time Pcr ◽  
The Difference ◽  
Estimated Breeding Values

Feed efficiency is an economically important trait in livestock and residual feed intake (RFI) is a commonly used measure of the trait in beef cattle. Residual feed intake is the difference between the actual feed intake recorded over a test period and the expected feed intake of an animal based on its size and growth rate. It is a heritable trait, and efficient animals have lower RFI values. Several genes have been identified as being differentially expressed in the liver of Angus bulls that have been divergently selected for RFI. The objective of this study was to use genes that are differentially expressed in bulls to classify Angus steers from the same divergent RFI selection lines. Liver samples were collected at slaughter from 40 high RFI and 40 low RFI steers that were ~23 months old, and had just completed a 251-day feedlot feeding period. RNA samples from the livers were assayed by quantitative real-time PCR for 14 genes, which have been identified previously in bulls. Steers were not measured for RFI, hence the estimated breeding values (EBV) for RFI of their parents were used to calculate their mid-parent (average of the two parents) RFI-EBV. Correlation and discriminant analyses were conducted on the normalised quantitative real-time PCR data from the steers. Discriminant analysis was also conducted on the bull data for comparison. In the steers, 8 out of the 14 genes were significantly (P < 0.05) correlated with RFI-EBV. Two genes from the glutathione S-transferase mu family (GSTM1 and GSTM2) and the S100 calcium-binding protein A10 (S100A10) had the highest correlations with RFI-EBV, with correlation coefficients of 0.59, 0.44 and 0.36, respectively. Based on the 14 expressed genes, 84% of the steers and 98% of the bulls were correctly classified into their respective RFI selection lines. The results of this study indicate that a high proportion of the genes that were differentially expressed in the original study with bulls were also differentially expressed in this study with steers. The high accuracy in classification obtained in this study shows that the transcriptional approach to the study of the biological processes involved in variation in RFI has great potential for identification of candidate genes.

scholarly journals Genetic polymorphism of CYP1A1 and CYP2D6 in populations of Buryats, Teleuts and Russians of Eastern Siberia

2018 ◽  
Vol 22 (2) ◽  
pp. 205-211 ◽  
Author(s):  
L. E. Tabikhanova ◽  
L. P. Osipova ◽  
T. V. Churkina ◽  
E. N. Voronina ◽  
M. L. Filipenko
Keyword(s):  
High Frequency ◽  
Genetic Research ◽  
Indigenous Populations ◽  
Eastern Siberia ◽  
Frequency Range ◽  
East Siberia ◽  
Gene Frequencies ◽  
Allele Specific ◽  
Polymorphic Variants ◽  
Frequency Occurrence

The study of the gene polymorphism of the system of biotransformation of xenobiotics is an important area of modern medical and genetic research. The aim of this work is to study the frequency of the alleles of the CYP1A1 (A2455G (*2C), rs1048943), CYP2D6 (A2549del (*3), rs35742686); G1846A (*4), rs3892097) genes of Teleuts (n = 115), Eastern Buryats (n = 132), Western Buryats (n = 280), their Métis (n = 56), and Russians of East Siberia (n = 122). Genotyping was performed using real-time PCR with competitive TaqMan allele-specific probes. The frequency of the CYP1A1*2C (2455G) allele was 28.8 % in the Eastern Buryat, 34.6 % in the Western Buryat, 16.7 % in the Teleut, and 31.3 % in the Métis cohort. The frequency of CYP1A1*2C (2455G) in the Russians of Eastern Siberia (4.1 %) corresponds to the frequency range found in European populations. A high-frequency occurrence of CYP1A1*2C (2455G) among Buryats and Teleuts may be indicative of a higher population-wide risk of diseases influenced by technogenic pollutants – substrates of CYP1A1. The CYP2D6*3 (2549del) allele was not detected in cohorts of indigenous populations, among Russians it was 0.4 %, and it was 2.7 % among Métis. The frequency of CYP2D6*4 (1846A) in Eastern and Western Buryats was 5.3 % and 4.3 %, respectively, for Teleuts it was 7.4 %. It was significantly higher in the Russian population (12 %), and among Métis (9.8 %). The obtained data makes it possible to predict a reduced risk of side effects of drugs and cancer associated with CYP2D6*3 (2549del) and CYP2D6*4 (1846A) in the Buryat and Teleut populations. However, metisation introduces new polymorphic variants into indigenous populations, shifts gene frequencies and changes the degree of risks.

Role of WNT4, HOXA10 and TWIST1 genes in the pathogenesis of external genital endometriosis and uterine leiomyoma

2021 ◽  
Vol 70 (3) ◽  
pp. 31-40
Author(s):  
Olga V. Malysheva ◽  
Arseny S. Molotkov ◽  
Natalya S. Osinovskaya ◽  
Natalya Yu. Shved ◽  
Maria I. Yarmolinskaya ◽  
...  
Keyword(s):  
Menstrual Cycle ◽  
Real Time ◽  
Real Time Pcr ◽  
Uterine Leiomyoma ◽  
Comparison Group ◽  
Reproductive Age ◽  
Cycle Phase ◽  
Transcription Analysis ◽  
Polymorphic Variants

BACKGROUND: Uterine leiomyoma and endometriosis are the most common gynecological diseases in women of reproductive age. A number of data indicate that there are common elements in the pathogenesis of these hyperproliferative conditions. This article is devoted to comparative analysis of the role of the WNT4, HOXA10 and TWIST1 genes in the development of uterine leiomyoma and external genital endometriosis. AIM: The aim of this study was to evaluate the frequency of polymorphic variants rs7521902 (WNT4) and rs4721745 (TWIST1) in patients with uterine leiomyoma, external genital endometriosis and in the comparison group; to determine the frequency of rare allelic variants of the HOXA10 gene in patients with external genital endometriosis; and to study the expression of these genes in the endometrium in patients with uterine leiomyoma, EGE and in the comparison group. MATERIALS AND METHODS: The polymorphic variants of the WNT4 and TWIST1 genes were studied by real-time PCR in patients with external genital endometriosis, uterine leiomyoma and in the comparison group. In patients with EGE and women in the comparison group, the second exon of the HOXA10 gene was sequenced. Real-time PCR with reverse transcription analysis of the expression of the WNT4, TWIST1 and HOXA10 genes in endometrial samples from the patients of the study groups was performed. RESULTS: The frequencies of polymorphic variants rs7521902 (WNT4) and rs4721745 (TWIST1) in patients with uterine leiomyoma, external genital endometriosis and in the comparison group did not differ significantly. Minor alleles of the HOXA10 gene were not identified in patients with external genital endometriosis. Expression of the WNT4 gene in the endometrium of patients with external genital endometriosis was independent of menstrual cycle phase and was reduced by 1.9 times compared to the endometrium of women with uterine leiomyoma. Expression of the HOXA10 gene in the endometrium of endometriosis patients on days 20-23 of the menstrual cycle was significantly reduced compared to the women in the comparison group. Expression of the TWIST1 gene was not altered in the endometrium of patients with uterine leiomyoma and external genital endometriosis. CONCLUSIONS: We did not identify associations of the studied polymorphic variants of the WNT4 and TWIST genes and minor variants of the HOXA10 gene with uterine leiomyoma and external genital endometriosis. The expression of the WNT4 and HOXA10 genes is reduced in the endometrium in patients with external genital endometriosis, but not in women with uterine leiomyoma. Changes in expression patterns of the studied genes in the endometrium differ significantly in these two diseases.

Design of a New Universal Real-Time PCR System Targeting the tuf Gene for the Enumeration of Bacterial Counts in Food

2010 ◽  
Vol 73 (4) ◽  
pp. 670-679 ◽  
Author(s):  
YUICHIRO TANAKA ◽  
HAJIME TAKAHASHI ◽  
USIO SIMIDU ◽  
BON KIMURA
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Real Time Pcr ◽  
Bacterial Species ◽  
Linear Regression Analysis ◽  
Bacterial Counts ◽  
Real Time Quantitative Pcr ◽  
Tuf Gene ◽  
Wide Range ◽  
The Difference

A novel universal real-time PCR, consisting of newly designed oligonucleotide subsets, was designed for a bacterial housekeeping gene encoding the peptide elongation factor Tu. Specificity and universality were confirmed in 66 bacterial strains, including 51 genera and 63 species. The amplification kinetics of tuf gene–targeted real-time quantitative PCR were consistent in a wide range of bacterial species tested. A calibration curve (r2 = 0.97) was produced for the estimation of bacterial counts, based on measurements of representative inoculations with 10-fold serial dilutions of the cells of representative bacterial species. Linear regression analysis of the real-time PCR–derived bacterial counts and aerobic plate counts, in a total 149 samples consisting of 25 minced meat, 34 fresh-cut vegetables, and 90 fish, exhibited a high correlation (r2 = 0.84, 0.87, and 0.95, respectively) over the range of 3.0 to 9.0 log CFU/g. In total, the difference between the two methods was less than 0.5 log in 75 of these samples, and in the remaining 74 samples, the difference was 0.5 to 1.0 log. Presently, our tuf gene–targeted real-time quantitative PCR assay achieves a rapid (within 2 h) estimation of bacterial counts of 3.0 to 9.0 log CFU/g, in a practical manner.

scholarly journals Evaluating Expression of Gene Encoding Flavonol Synthase in Green TrungDu and Purple TrungDu by Real time PCR and HPLC Technique

2020 ◽  
Vol 36 (2) ◽  
Author(s):  
Hoang Thi Thu Yen ◽  
Nguyen Thuy Linh ◽  
Huynh Thi Thu Hue ◽  
Nguyen Hai Dang
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Expression Level ◽  
Gene Encoding ◽  
Flavonol Synthase ◽  
Apical Bud ◽  
Young Leaves ◽  
Hplc Technique ◽  
The Difference ◽  
The Relationship

In plants, flavonol synthase (FLS) is a multifunctional enzyme that converts dihydroflavonol into flavonols and naringenin into dihydrokaempferol. FLS from tea has been shown to metabolize dihydroquercetin to quercetin. In this study, we conducted studying on the relationship between quercetin content and the expression level of FLS in two traditional tea cultivars of Vietnam, TrungDuxanh and TrungDutim tea grown in tea garden of Thai Nguyen University of Agriculture and Forestry. Tea shoots with one apical bud and two to three young leaves using as research materials which collected in September 2017. The using of HPLC technique did not detect the quercetin content from the these tea samples. The preliminary results of quantification of FLS gene expression by real time PCR showed the expression level of FLS in green Trung Du tea is higher than purple Trung Du, although the difference is not great. Thus, at the time of collecting, the expression of FLS in green Trung Du and purple Trung Du can not synthesize quercetin but synthesize other flavonols.

scholarly journals QUANTIFICATION OF STREPTOCOCCUS SANGUINIS ISOLATED FROM DENTAL PLAQUE AND SALIVA OF SUBJECTS WITH AND WITHOUT CORONARY HEART DISEASE – ANALYSIS USING REAL-TIME PCR

2017 ◽  
Vol 20 (1) ◽  
pp. 13-19
Author(s):  
Aini Hariyani Nasution ◽  
Yulianti Kemal ◽  
Robert Lessang ◽  
Boy Bachtiar
Keyword(s):  
Coronary Heart Disease ◽  
Heart Disease ◽  
Real Time ◽  
Real Time Pcr ◽  
Dental Plaque ◽  
Vascular Diseases ◽  
Specific Primers ◽  
Streptococcus Sanguinis ◽  
The Difference ◽  
Disease Analysis

Coronary Heart Disease (CHD) is the major cause of death in most countries in the world. Gram-positive and Gram-negative bacteria have been identified in bacteremia cases and known to have a role in various vascular diseases, including Streptococcus sanguinis which is most frequently isolated from endocarditis patients and often associated with CHD. The purpose of this study was to analyze the number of Streptococcus sanguinis isolated from dental plaque and saliva of subjects with and without CHD. Bacterial colonies isolated from the dental plaque and saliva of 16 subjects without CHD and 8 subjects with CHD were planted in Mitis salivarius agar, and then the DNA was extracted and quantified with a Real-Time PCR technique using 16S rRNA specific primers. The quantification of Real-Time PCR showed that there was a difference in the number of S. sanguinis between the two groups of subjects, but an unpaired T-test showed that the difference was not statistically significant. Furthermore, the number of S. sanguinis from dental plaque in CHD subjects tends to be higher than that of non-CHD subjects whereas the number of S. sanguinis from saliva in non-CHD subjects tends to be higher than that of CHD subjects.

scholarly journals TaqMan-Based Real-Time PCR for Genotyping Common Polymorphisms of Haptoglobin (HP1 and HP2)

2008 ◽  
Vol 54 (11) ◽  
pp. 1908-1913 ◽  
Author(s):  
Mikiko Soejima ◽  
Yoshiro Koda
Keyword(s):  
Real Time ◽  
Internal Control ◽  
Real Time Pcr ◽  
Relative Number ◽  
High Throughput Analysis ◽  
Large Populations ◽  
Allele Specific ◽  
Taqman Pcr ◽  
Pcr Method ◽  
The Difference

Abstract Background: The haptoglobin gene (HP) has 2 common codominant alleles (HP1 and HP2) that account for 3 phenotypes. HP2 is generated by a 1.7-kb intragenic duplication of HP1. Methods: We used the real-time TaqMan PCR system to develop an effective method for HP genotyping that allows us to evaluate the relative number of copies of the HP2 allele–specific junctional region of the 1.7-kb gene duplication (HP2) by comparing the intensity of the amplification signals to those of the HP promoter region (HP5′), which was used as the internal control. The difference in threshold cycles (ΔCt) between HP2 and HP5′ was used to assess HP2 copy number. In addition, the assay detects the HP deletion (HPdel) at the same time. Results: The mean 2−ΔΔCt values (the HP2/HP5′ ratio) obtained from 123 samples of known HP genotypes clearly differentiated 2 nonoverlapping intervals that correspond to the HP genotypes. Ratios for HP2/HP1 samples ranged from 0.34–0.50, HP2/HP2 samples ranged from 0.79–0.98, and the absence of an HP2 allele signal was defined as HP1/HP1. We simultaneously detected HPdel. The assay produces results in &lt;1 h. Conclusions: The TaqMan-based real-time PCR method was successfully applied to HP genotyping. The method is easy to use in a molecular diagnosis laboratory, and its robustness and rapidity make it suitable for high-throughput analysis of large populations.

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.

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