Hybridization-fluorescence Detection Research Articles

ANALYSIS OF THE SPECIES COMPOSITION AND INFECTION OF TICKS WITH PATHOGENS OF LYME-DISEASE IN THE TERRITORY OF THE REPUBLIC OF TATARSTAN FOR THE PERIOD 2012 - 2022.

Rationale: Analysis of the manifestations of the epizootic process is important for planning preventive and anti-epidemic measures and is an integral component of epidemiological surveillance of natural focal infections. Purpose of the study: To analyze the species composition of ticks and the infection of ticks with borreliosis pathogens in the Republic of Tatarstan (RT) for the period 2012 - 2022. Methods: An analysis of the species composition of ticks collected by the zoological group of the Center for Hygiene and Epidemiology in the RT from environmental objects during the period 2012-2022 was carried out. Testing of the ticks for B. burgdorferi and B. miyamotoi was carried out using the polymerase chain reaction method with hybridization-fluorescent detection. Results: The total number of people who applied for tick bites in the Republic of Tatarstan in 2013 – 2022 was 77,703. During the study period, 5536 ticks were collected for research on the territory of the Republic of Tatarstan from environmental objects. The species composition of ticks: Dermacentor reticulatus – 75.4%, Ixodes ricinus – 14.1% and Ixodes persulcatus – 10.5%. Infection of different types of ticks with Borrelia was not the same and was 20.4% for Ixodes persulcatus, 14% for Ixodes ricinus, 4.4% for Dermacentor reticulatus. In 4 regions of the republic, from ticks of the species Ixodes ricinus pathogenic genomospecies Borrelia miyamotoi were foumd; (the infestation of this type of tick with Borrelia miyamotoi was 0.4%). Indicators of morbidity of the population with ixodid tick-borne borreliosis (TBB) and indicators of infection rates for ticks vary in different regions of the republic. Conclusion: The dominant species of ixodid ticks in natural epitopes of the Republic of Tatarstan in 2012-2022 were Dermacentor reticulatus (75.4% of the total number of collected ticks). For ticks of this species, the lowest level of infection with Borrelia was demonstrated (4.4% of ticks). This may be one of the factors responsible for the low incidence rates of tick-borreliosis in the Republic of Tatarstan. The identification of ticks of the species I. ricinus infected with Borrelia miyamotoi confirms the need for targeted molecular genetic and serological examination for recurrent fever in patients with non-erythematous forms of febrile illness after an episode of tick bite.

ТЕСТ-СИСТЕМА ДЛЯ ПРОВЕДЕНИЯ ПЦР В РЕЖИМЕ РЕАЛЬНОГО ВРЕМЕНИ ДЛЯ ОБНАРУЖЕНИЯ ДНК ТТV В БИОЛОГИЧЕСКОМ МАТЕРИАЛЕ

Background. The study of biological material for the presence of TTV DNA using the PCR method allows for a timely assessment of the functional state of the human liver and immune system. Objective. To develop components for real-time PCR for TTV DNA detection in biological material. Material and methods. The design and selection of optimal primers and probes (taking into account the size (length) of the amplicon, annealing temperature, nucleotide composition, distribution of nucleotides along the length of the primer, length of primers, the possibility of formation of hairpins and dimers by primers) were performed using the Primer-BLAST/Primer3, FastPCR programs. Since primers, even absolutely unique for certain DNA sequences, could anneal at nonspecific sites, not related to the gene analyzed, we checked the correspondence of the primers to the sequences of the target gene. For this purpose, we used the NCBI Primer BLAST online service and assessed the local pairwise alignment of each primer with all nucleotide sequences of the Refseq databases. Results. As the result of studies carried out on the selection of the optimal primer annealing temperature, primer concentrations, as well as the selection of the optimal nucleotide pair, the main parameters of the designed primers were determined. Conclusions. A kit for the detection and quantification of TTV DNA using the polymerase chain reaction method with hybridization-fluorescent detection in real time was created and became the basis for the development of a commercial test system.

Биохимический состав семян сои Дальневосточного региона

The purpose of the study is to analyze seeds of cultivated and wild soybeans in the Far Eastern Region according to a number of indicators of biochemical composition. The object of the study is seeds of 18 varieties and 4 forms of wild soybeans grown in the Amur Region (Alena, Lidiya, Kitrossa, Sonata, Garmoniya, KBL-29, KA-1413, KZ-6337), Khabarovsk (Marinata, Batya, VAZ-100, Ivan Karamanov, Saltus, wild soybean HK), Primorsky Regions (Musson, Primorskaya -96, Primorskaya-4, Primorskaya-86, Sfera) and the the Democratic People's Republic of Korea (black, yellow, green). The content of transgenes in seeds was determined by PCR with hybridization-fluorescent detection in real time (Real Time PCR); concentration of protein, oil and VVC – by the method of diffuse reflection in the near infrared region. The graphs obtained as a result of amplification indicate that during the PCR analysis, no gene-tically modified construct (35S promoter and NOS terminator) was detected in any of the soybean varieties studied. The studied soybean varieties from the Far Eastern Region are not transgenic. According to biochemical indicators, the Kitrossa variety of Amur selection should be distinguished, which is characterized by a high content of protein (46.80 ± 0.02 %), oleic (27.53 ± 0.01) and linolenic acids (11.79 ± 0.01 %), as well as the DPRK soybean varieties (from 41.43 ± 0.03 to 43.46 ± 0.01 %), which had an increased protein content with an optimal linoleic-linolenic ratio (ω-6: ω-3). Wild soybeans have been shown to contain high amounts of protein. For the KZ-6337 form, the maximum amount of protein was identified (50.06 ± 0.10 %), therefore it is recommended to use it as a source of dominant genes when breeding high-protein soybean varieties. The results of a study of the biochemical composition of soybeans in the Far Eastern Region indicate the varietal specificity of the region of growth in protein, oilseed and fatty acid composition.

Prevalence of occult hepatitis B infection among blood donors in Saint Petersburg

The aim of this study was to assess the prevalence of occult hepatitis B infection among blood donors in St. Petersburg, as well as to characterize the identified virus isolates. The study material was represented by 2800 blood plasma samples collected in 2019 from blood donors living in St. Petersburg. The ELISA study for HBV marker rate consisted of HBsAg, anti-HBs IgG, anti-HBcore IgG. HBV DNA was analyzed by nested PCR with real-time hybridization-fluorescence detection on three targets allowing to determine virus DNA at low viral load, including HBsAg-negative chronic hepatitis B. Hepatitis B serological markers were detected in 69.43% of those surveyed, HBsAg was found in 0.43% of individuals, and all of which donated blood first time. A significant excess of the anti-HBcore IgG antibodies occurrence among primary donors (15.1%) compared with repeated/regular donors (7.48%) was shown. The prevalence of virus DNA in the group was 3.14%, including 2.71% of cases in HBsAg-negative CHB. Based on phylogenetic analysis of 88 isolates, HBV subgenotypes were determined in the following order: D1 and D2, 40.91% each, D3 and A2, 9.09% each. While determining the serological subtype in detected isolates, the serotype ayw3 (52.27%) vs ayw2 (46.59%) and adw2 (10.23%) prevailed. Drug resistance mutations, including compensatory ones, were detected in six examined patients (6.82%). In all genotype D isolates, multiple amino acid substitutions were identified in the RT, SHB, MHB, LHB, and Core regions; mutations in the preCore region were detected in 21.59% samples. In the MHR of the HBV genotype D genome, twenty-six positions were identified in which amino acid substitutions occurred, and all isolates showed modifications at positions 113, 114, 131, 134, 159, 161, 168, in 76 — at position 122, in 68 — at position 127, in 36 — at position 118, in 24 — at position 128. In HBV A2 isolates, mutations T113S, S143T, Y161F were identified. Nine isolates in the preCore region showed a polymorphism including a stop codon W28*W; in five isolates the W28S substitution was shown in the same position, and the W28*S variant was found in one more sample. The high incidence of HBsAg-negative CHB cases among blood donors, as well as the predominance of HBV isolates that simultaneously carry mutations resulting in diagnostic failure of HBsAg tests and prophylactic failure of immunoglobulin or vaccines and virus reactivation, mutations that contribute to disease progression obviously pose a threat to health and require to be further examined.

The structure of some blood-borne infections distribution among persons from penitentiary institutions the markers

The aim of the study was to assess the occurrence of serological and molecular genetic markers of HIV infection, viral hepatitis B and C among persons in pre-trial detention.Materials and methods. The material of the study identified 138 samples of blood sequences obtained from men in the pretrial detention center in St. Petersburg. Patients were examined by ELISA for the presence of serological markers of HIV infection (Ag/Ab HIV), HBV (HBsAg, antiHBs IgG, anti-HBcore IgG) and HCV (anti-HCV IgG). Determination of HIV and HCV RNA, as well as HBV DNA by PCR with hybridization-fluorescence detection in «real time» mode. HBV DNA with a low viral form was detected using a technique developed at the Saint-Petersburg Pasteur Institute.Results and discussion. With the prevalence of the general prevalence of serological markers, markers of HIV infection were detected in 55 (39.86%) patients, HBV — in 73 (52.90%), HCV — in 83 (60.14%). the same markers of HIV infection and HBV were detected in 6 (4.35%) examined, HIV infection and HCV in 17 (12.32%), HBV and HCV in 22 (15.94%) persons, to all three infections in 26 (18.84%) cases. In 22 (15.94%) people did not notice any of the analyzed markers. HIV RNA was found in 23.19%, HCV RNA — in 46.38%, HBV DNA — in 10.14% of the examined. At the same time, HBV DNA with a low viral form (less than 50 IU/ml) was detected in 7.97% of cases, including 6.52% of seronegative samples. Simultaneously, HIV and HCV RNA were detected in 6.52%, HCV RNA and HBV DNA in 2.17% of patients. Molecular biological markers of all three indicators were found in 1.45% of patients. The examined group was ranked by age into the following subgroups: 21– 30 years old (12.32%), 31–40 years old (34.78%), 41–50 years old (37.68%) and older than 51 years old (15.22%). Certain differences in the prevalence and distribution of the analysis of serological and molecular biological markers between age markers, the highest frequency was found for groups of 31–40 and 41–50 years.Conclusion. The results obtained indicate a high prevalence of HIV infections and parenteral viral hepatitis B and C among persons convicted and awaiting a stage in a pre-trial detention center. Careful attention must be paid to their admission to penitentiary institutions, since many convicts are not aware of their morbidity and may become infected. Regular conclusions, concluded in dynamics, including laboratory diagnostics using highly sensitive molecular biological methods, upon admission to a penitentiary institution, and then annually and upon release, should be taken into account as measures to prevent infection with HIV, HBV, HCV.

Development of a Reagent Kit for the Quantitative Determination of Hepatitis B Virus (HBV) DNA in Clinical Material by PCR with Hybridization-Fluorescence Detection

Relevance.Hepatitis B virus (HBV) is one of the most common viral infections affecting people worldwide and can lead to chronic hepatitis, cirrhosis and hepatocellular carcinoma. Currently 3% of the world's population are infected with hepatitis B virus and are at risk of developing life-threatening liver disease. Immunological and molecular biological methods of detection of HBV are currently used in laboratory diagnostics. The polymerase chain reaction (PCR) is currently the most sensitive method for the detection and quantification of HBV. HBV DNA quantification is widely used to monitor the antiviral treatment of HBV infection.Aim.To develop a real-time PCR kit for the quantification of HBV DNA.Materials and methods.A total of 200 plasma and serum samples positive and negative for HBV were used in the development. The performance of the developed kit was compared with the use of other commercially registered HBV diagnostic kits in Russia. Additionally, the nucleotide sequences of all existing virus genotypes analysed for the selection of primers using GeneBank system.Results and discussion.Comparison analysis of the results of quantitative determination by real-time PCR in 200 clinical serum and blood plasma samples showed that the diagnostic sensitivity of the developed kit was 100% and specificity 100%. The primers developed specific to the POL gene region. The kit is capable of detecting all types of virus genotypes.Conclusions.The developed reagent kit allows detection of hepatitis B virus and determination of its quantity within 70 minutes. In addition to a large number of genotypes and subgenotypes, the virus is characterized by mutational changes in the genome, which complicates its diagnosis and, as a consequence, the ongoing therapy with drugs. Conservative regions for primer and probe selection taken into account in the development, and the sequencing results obtained are applicable to all HBV genotypes. The reagent kit is designed to monitor HBV infected patients and will allow the analysis of different HBV viral loads.

Prevalence of HIV and Viral Hepatitis Markers among Healthcare Workers in the Republic of Guinea.

Healthcare workers are much more likely to be infected with HIV and hepatitis viruses compared to the general population. Although healthcare workers are more aware of HIV and hepatitis viruses, several countries in Africa lack a comprehensive grasp of disease routes and transmission risks. The aim of this study was to assess the prevalence of the serological and molecular biological markers of HIV and viral hepatitis among healthcare workers in the Republic of Guinea. The study material was 74 blood serum samples collected from healthcare workers who received additional training at the Institute of Applied Biological Research of Guinea (IRBAG, Kindia, Republic of Guinea). The markers examined included HBsAg, HBeAg, anti-HBs IgG, anti-HBcore IgG, anti-HCV qualitative determination, anti-HEV IgM and IgG, anti-HAV IgM and IgG, and anti-HIV. For viral DNA and RNA detection, nucleic acids were extracted from blood serum, and viral presence was inferred using real-time PCR with hybridization fluorescence detection. A high prevalence of viral hepatitis B markers was shown, and significantly fewer cases of viral hepatitis C and HIV were detected. Almost all examined medical workers had anti-HAV IgG antibodies, but no antibodies to hepatitis E virus. Apparently, the identified markers depend on the general prevalence of certain pathogens in the region and are associated with the traditions and characteristics of the country's residents.

Выявление бактерий рода Campylobacter в используемой для искусственного осеменения сперме быков

The article presents the results of molecular genetic studies of samples of cryopreserved bovine semen used for artificial insemination from Russian and foreign breeding farms, aimed at identifying the genetic material of Campylobacter of different species. Studies on the detection of C. jejuni DNA were carried out by PCR with electrophoretic detection of PCR products using the KAM-BAK test system, Campylobacter spp. and C. fetus – using LSI VetMAX™ PCR kits with realtime hybridization-fluorescence detection. The genetic material of microorganisms of the genus Campylobacter was detected by PCR in 66,9 % of the studied samples, C. fetus – in 0,7 %, C. jejuni – in 9,7 % of semen samples. For 34 randomly selected samples of semen, a study of the microbial composition was carried out using «next generation» sequencing technology (NGS). Campylobacter has been shown to be the third most common genus of bacteria in semen samples. A species of C. ureolyticus that causes gastrointestinal diseases in humans and was previously found in feces and milk of cattle has been identified in semen samples. The issues of penetration of different types of Campylobacter into semen and the impact of the presence of Campylobacter on the quality of sperm production are discussed.

Development and study of the performance of a reagent kit for the identification of <i>Vibrio cholerae</i> and <i>Vibrio parahaemolyticus</i> by multilocus allele-specific polymerase chain reaction in clinical trials

BACKGROUND: Cholera belongs to the group of infections, and the fight against it is regulated by the WHO International Health Regulations (2005). To date, several test systems have been developed for the polymerase chain reaction (PCR) detection of Vibrio cholerae strains. However, no PCR test systems have been registered for detecting V. parahaemolyticus DNA in clinical material in Russia.
 AIM: To conduct clinical trials and evaluate the performance of the Reagent kit for identification of V. cholerae and V. parahaemolyticus by multilocus allele-specific PCR (kit of reagents Vibrio-screen FL) for the state registration of the specified medical device.
 MATERIALS AND METHODS: The study presents the results of the clinical trials for the reagent kit Vibrio-screen FL that has been developed to optimize the means and methods of identification and differentiation of pathogens of dangerous infections based on data on the diversity of genomes. Clinical trials were conducted at the Head Center of Hygiene and Epidemiology of the FMBA of Russia (license from 02.12.2013 No. ФС-99-01-008342; permission of the Ministry of Health from 16.05.2013 No. 300n) in accordance with the program of clinical trials No. 20.ПМКИ-042/10.20 from 23.09.2020 (Clinical Trial Certificate No. 01.AK-042/10.20 dated 12.10.2020). The clinical trials of the developed reagent kit Vibrio-screen FL used clinical strains of microorganisms of the genus Vibrio and other microorganism species as target analytes. Bacteriological method and multilocus PCR with hybridization-fluorescence detection were used.
 RESULTS: The reagent kit Vibrio-screen FL is a medical device for professional use in clinical laboratory diagnostics. The analytical sensitivity of the reagent kit Vibrio-screen FL was at least 106 copies of the SecY gene when determining belonging to the genus Vibrio; to the species V. cholerae, at least 106 copies of the hlyA gene; and to the species V. parahaemolyticus, at least 106 copies of the vppC gene. When determining the analytical specificity, no non-specific reactions with DNA samples of other microorganism species were found. A medical device for in vitro diagnostics A reagent kit for the identification of V. cholerae and V. parahaemolyticus by multilocus allele-specific PCR (Vibrio-screen FL) meets the requirements of the intended use, which allows it to be used for the differentiation of V. cholerae and V. parahaemolyticus strains isolated from a cultured clinical samples by multilocus allele-specific polymerase chain reaction with hybridization-fluorescent detection. The registration certificate No. РЗН 2021/13360 from 08.02.2021 was received. By the order of Roszdravnadzordated 08.02.2021 No. 1041, a reagent kit Vibrio-screen FL by the Rostov-on-Don Anti-Plague Institute of Rospotrebnadzor was admitted to distribution on the territory of the Russian Federation.
 CONCLUSIONS: The reagent kit Vibrio-screen FL is a high-quality, effective, and safe diagnostic medical device for in vitro diagnostics with sufficient diagnostic sensitivity and specificity to identify specific fragments of Vibrio spp., V. cholerae, and V. parahaemolyticus nucleic acids. These allows it to be used in clinical laboratory diagnostics for epidemiological analysis and registration of diseases caused by pathogenic vibrions.

The structure of intestinal infections in the infectious department

Background. Every year in Russia, the number of patients with acute intestinal infections of unknown etiology remains at a significant level.
 Aim. To determine the current etiological structure and features of the clinical course of acute intestinal infections in the infectious diseases department of a city hospital.
 Material and methods. For the period from 2017 to 2019, 181 patients with manifestations of acute intestinal infections, who were in the infectious departments of the City Hospital No. 5, Barnaul, were examined. The age of the patients ranged from 18 to 76 years (mean age 35.414.8 years), including 55.6% females and 44.4% males. The study of biological material from patients (feces) was performed using polymerase chain reaction with hybridization-fluorescence detection, bacteriological examination and serological method (indirect hemagglutination reaction) for the presence of antibodies to pathogens of acute intestinal infections (in blood serum). The statistical significance of differences was assessed by the 2 test.
 Results. Out of 181 patients with acute intestinal infections, 108 (59.7%) had the etiology of the disease, of which 54 (50%) patients had an infection of viral etiology, including in the form of monoinfection in 45 cases, and caused by a combination of two viruses in 9 cases. In 41 (38%) patients, acute intestinal infections of bacterial etiology were diagnosed, including a combination of two pathogens in 4 cases, and three pathogens in 1 case. 13 patients had viral-bacterial intestinal infections. As a result of the survey, on the existing list of laboratory tests, the etiology of intestinal infection was not established in 73 (40.3%) patients. In patients with acute intestinal infection of bacterial etiology, the disease proceeded more severely, with higher body temperature and colitis, in patients with intestinal infection of viral etiology with gastroenteritis.
 Conclusion. A high proportion of intestinal infections of viral etiology (up to 50%) with etiological dominance of noroviruses has been established. Among the causative agents of bacterial acute intestinal infections, Campylobacter and Salmonella prevailed.

Role of human papillomavirus infection in the development of oral leukoplakia and oral squamous cell carcinoma

BACKGROUND: The annual global oral cancer morbidity is more than 650,000 cases. Between 2007 and 2017 in the Russian Federation, the absolute number of first-ever diagnosed oral cavity malignant neoplasms is constantly increasing. According to many authors, oral squamous cell carcinoma is associated with pre-existing leukoplakia. This disease is the most common and prone to malignant precancerous oral mucosa disease. Human papillomaviruses (HPV) are crucial players in this process. However, the relationship between HPV infection in leukoplakia and oral squamous cell carcinoma remains unclear.
 AIM: To establish the HPV role in the development of leukoplakia and squamous cell carcinoma of the oral mucosa.
 MATERIAL AND METHODS: The study included 39 patients with verified various forms of oral leukoplakia and oral squamous cell carcinomas. The squamous cell carcinoma group included 19 patients. The oral leukoplakia group is consisted of 20 patients. The control group of healthy volunteers is represented by 22 patients with clinically healthy oral mucosa. HPV deoxyribonucleic acid (DNA) was detected and quantified in clinical materials by polymerase chain reaction with hybridization-fluorescence detection AmpliSense HPV HRS screen-titer-FL for HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68 and AmpliSense HPV 6/11-FL for HPV types 6 and 11.
 RESULTS: HPV types 6 and 11 DNA were isolated in three cases with insignificant clinical disease from 19 samples taken from patients with oral squamous cell carcinoma. In groups of leukoplakia and healthy volunteers, no high oncological risk was identified. Despite a HPV-positive expression in squamous cell carcinoma samples is detected, no significant difference was found between the squamous cell carcinoma group and control group (p=0.09).
 CONCLUSION: No significant association was noted between HPV and oral squamous cell carcinoma; however, a low carcinogenic-risk HPV was detected in 15.7% of carcinoma samples. No HPV contamination was detected in patients with leukoplakia and healthy groups.

Identification of genetically modified organisms in foods of plant origin as a way to control health risks for consumers

Uncontrolled distribution of goods produced by genetically modified plants should be prevented by the state in order to secure food safety in the Russian Federation and to minimize health risks for consumers. We analyzed foods of plant origin for children to identify components of genetically modified organisms in them. It was done to ensure safety of such foods. The highest specific weight among the analyzed foods belonged to nectars (40.0 %) and juice-containing drinks (36.0 %). Juices and fruit drinks accounted for 12 % each. Genetically modified organisms were determined in foods by identifying regulatory sequences (35S promoter, FMV promoter and NOS terminator) that are widely used in constructions of genetically modified plants. Occurrence of regulatory genetic elements specific for genetically modified organisms was checked in juice products for children by the polymerase chain reaction in real-time mode with hybridization-fluorescent detection of amplification products and with the use of the “AmpliSens GM Plant-1-FL” and “AmpliKvant GM soya-FL” test systems. The results of this study showed that no analyzed foods of plant origin contained any regulatory sequences (35S, NOS и FMV) indicating presence of genetically modified organisms. Fluorescence through the FAM, Cy5 and ROX channels did not exceed its threshold value. Therefore, we did not detect any violations of the established requirements to occurrence of genetically modified organisms in foods for children. Further investigation that would involve examining a more extensive material is required to ensure proper assessment and control of food contamination with genetically modified organisms in order to ensure food safety.

Идентификация генетически модифицированных организмов в пищевой продукции растительного происхождения как средство контроля рисков для здоровья потребителей

Uncontrolled distribution of goods produced by genetically modified plants should be prevented by the state in order to secure food safety in the Russian Federation and to minimize health risks for consumers. We analyzed foods of plant origin for children to identify components of genetically modified organisms in them. It was done to ensure safety of such foods. The highest specific weight among the analyzed foods belonged to nectars (40.0 %) and juice-containing drinks (36.0 %). Juices and fruit drinks accounted for 12 % each. Genetically modified organisms were determined in foods by identifying regulatory sequences (35S promoter, FMV promoter and NOS terminator) that are widely used in constructions of genetically modified plants. Occurrence of regulatory genetic elements specific for genetically modified organisms was checked in juice products for children by the polymerase chain reaction in real-time mode with hybridization-fluorescent detection of amplification products and with the use of the “AmpliSens GM Plant-1-FL” and “AmpliKvant GM soya-FL” test systems. The results of this study showed that no analyzed foods of plant origin contained any regulatory sequences (35S, NOS и FMV) indicating presence of genetically modified organisms. Fluorescence through the FAM, Cy5 and ROX channels did not exceed its threshold value. Therefore, we did not detect any violations of the established requirements to occurrence of genetically modified organisms in foods for children. Further investigation that would involve examining a more extensive material is required to ensure proper assessment and control of food contamination with genetically modified organisms in order to ensure food safety.

CHANGES IN THE COMPOSITION OF THE GUT MICROBIOTA IN THE OBESE HYPERTENSIVE PATIENTS

Objective: Recent studies showed that the gut microbiota (GM) is involved in the regulation of blood pressure and the pathogenesis of arterial hypertension (AH). Changes in the composition of the GM are associated with various metabolic diseases and the risk factors of development of AH, such as obesity and diabetes mellitus. The aim of this study was to investigate the features of the composition of the GM in the patients with AH, which occurs against the background of abdominal obesity (AO). Design and method: In the study was included 70 patients with AH, 2–3 degrees (46 patients with AO and 24 patients with normal body weight (NBW) aged 40 to 65 years. The control group consisted of 20 practically healthy individuals. The investigation complex included standard clinical, laboratory and instrumental methods. Determination of the quantitative composition of GM was carried out by the method of polymerase chain reaction with hybridization-fluorescence detection of results in real time. Results: In patients with AH, both with AO and without obesity (with NBW), there was a significant decrease in the number of Lactobacillus spp. (p < 0.05), Bifidobacterium spp (p < 0.05), Faecalibacterium prausnitzii (p < 0.05) compared with the control group/ At the same time, in the group of the patients with AH and AO, a more significant decrease in the number of Faecalibacterium prausnitzii was detected, compared with the group of patients with AH with NBW (p < 0.05). The amount of Akkermansia Muciniphila was significantly lower in the patients with AH and AO compare with the patients with NBW (p < 0.05) and the control group (p < 0.01). Also, in the patients with AH and AO the ratio of Bacteroides fragilis group/Faecalibacterium prausnitzii was significant higher than in the patients with NBW (p < 0.05) and control group (p < 0.05). Conclusions: The results of our study indicate a possible role of the identified GM imbalance (deficiency of Lactobacillus spp., Bifidobacterium spp, Faecalibacterium prausnitzii Akkermansia Muciniphila, increased ratio of Bacteroides fragilis group/Faecalibacterium prausnitzii) in the pathogenesis of AH in the obese patients.

DETECTION OF VIRUSES OF THE RUGOSE WOOD COMPLEX ON VINEYARDS OF THE ODESSA REGION

DETECTION OF VIRUSES OF THE RUGOSE WOOD COMPLEX ON VINEYARDS OF THE ODESSA REGION

ПЦР-диагностика возбудителей болезней сортов картофеля (Solanum tuberosum)

When creating technological schemes for the improvement of potatoes, PCR methods are currently actively used, in particular, amplification with hybridization-fluorescence detection in the "real time" and "end point" modes. In this work, both of these methods were applied and the diagnostics of cultivated potato varieties and lines for the presence of viral pathogens A, M, S, X, Y, Andean mottle viruses and latent potato thymovirus (APMV, APLV), as well as tuber spindle viroid (PSTVd) and potato panicle top virus (PMTV) was carried out. The analysis for the presence of parasitic diseases of pale Globodera pallida and golden cyst nematodes Globodera rostochiensis, bacterial ring and brown rot Clavibacter michiganensis subsp. sepedonicus and Ralstonia solanacearum, as well as fungal cancer Synchytrium endobioticum. As a result of PCR analysis, parasitic, fungal and bacterial diseases were not detected 44% of the total number of samples were healthy from viral diseases, while 25% were initially infected and underwent sanitation operations. For the first time, the PCR diagnostic system was introduced into the technological processes for obtaining varietal crops of the Tyumen region, free from diseases, to adapt to growing conditions in Western Siberia.

The features of the composition of the gut microbiota in patients with hypertension and abdominal obesity

The aim of this work was to study the features of the composition of the gut microbiota (GM) in patients with arterial hypertension (AH), which occurs against the background of abdominal obesity (AO). Materials and methods. The paper presents the data on examination of 70 patients with AH of 2–3 degrees (46 patients with AO and 24 patients without obesity – with normal body weight (NBW)). The control group included 20 apparently healthy individuals with NBW. The patients underwent detailed standard clinical, laboratory and instrumental examination. Determination of the quantitative composition of GM was carried out by the method of polymerase chain reaction with hybridization-fluorescence detection of results in real time using the test system “COLONOFLOR-16 (biocenosis)” (“ALFA-LAB”). In this work, a statistical analysis of the obtained data was carried out using standard methods and Microsoft Excel 17.0. Results. A significant change in the composition of GM were found in patients with AH, which proceeds against the background of AO, in comparison with apparently healthy individuals and with patients with AH without obesity (with NBW). In the group of AH patients with the presence of AO, significant decrease in the number of Lactobacillus spp., Bifidobacterium spp., Faecalibacterium prausnitzii and Аkkermansia muciniphila was found in comparison with apparently healthy individuals. The number of Faecalibacterium prausnitzii and Аkkermansia muciniphila in these patients was also significantly lower than in hypertensive patients with NBW. At the same time, the ratio of Bacteroides fragilis/Faecalibacterium prausnitzii was significantly higher than in the control group and the group of hypertensive patients without obesity – with NBW. Conclusions. The results of the study indicate a possible role for the deficiency of such representatives of GM as Lactobacillus spp., Bifidobacterium spp., Faecalibacterium prausnitzii and Аkkermansia muciniphila and an increase in the ratio of Bacteroides fragilis/Faecalibacterium prausnitzii in the pathogenesis of AH, which occurs against the background of AO.

Prevalence of hepatitis B and C viral markers among apparently healthy residents of the Socialist Republic of Vietnam (Southern Vietnam)

The aim of this study was to assess the prevalence of serological and molecular biological markers of viral hepatitis B and C among apparently healthy residents of the Southern Vietnam. The study material was represented by 397 blood serum samples collected from apparently healthy residents of the Southern Vietnam. The ELISA examination for presence of HBV and HCV markers involved HBsAg, anti-HBs IgG, anti-HBcore IgG, and anti-HCV qualitative determination. For HBV DNA and HCV RNA detection, nucleic acids were extracted from serum blood, and a test for virus detection was carried out by real-time PCR with hybridization fluorescence detection. Amplification and subsequent sequencing of HBV and HCV were performed using nested PCR with paired overlapping primers jointly flanking the target regions. Analysis of the overall prevalence of serological markers showed that among the apparently healthy individuals anti-HBsAg and anti-HCV antibodies were detected in 12.3% (95% CI: 9.27–15.99%) and 3.27% (95% CI: 1.76–5.53%) of individuals, respectively. The prevalence of HBsAg in men (19.1%) significantly exceeded that of found in women (5.9%), χ2 = 14.688 with p = 0.0001, df = 1, calculated odds ratio OR = 3.751 (95% CI: 1.892–7.439). Among apparently healthy patients, taking into account HBsAg-positive and negative samples, HBV DNA was detected in 26.95% (95% CI: 22.65–31.6%). HBV phylogenetic analysis showed that subtype B4 prevalence comprised 64.49%. Subtypes C1 (14.95%), B2 (9.35%), C2 (6.54%), C3 (0.93%), and C5 (3.74%) were also identified. HCV RNA was detected in 7 samples, which accounted for 1.76% (95% CI: 0.71–3.6%). Phylogenetic analysis showed that all HCV isolates belong to genotype 6, subtype 6a (100%).

Viral load as a factor of persistence of papillomavirus infection in cervical intraepithelial neoplasia grade I

Introduction. The viral antigenic load can influence the nature of the infectious response, leading to elimination of the virus or to chronicity of the process, and in some cases to the progressive course of the disease. The aim of the study was to investigate the relationship between the types of viral load of human papillomavirus and the age of a patient with cervical cervical intraepithelial neoplasia grade I. Material and methods. 86 patients with cervical papillomavirus-induced cervical intraepithelial neoplasia grade I by liquid oncocytology were examined. The frequency of human papillomavirus genotypes of high carcinogenic risk in this group of women was analyzed taking into account the viral load by polymerase chain reaction with real-time hybridization-fluorescence detection. The correlation between the viral load of the human papillomavirus and the age of the patient with further prediction of the regression or progression of cervical pathology was studied. Results. This study found that female patients under 30 years of age with cervical intraepithelial neoplasia grade I associated with papillomavirus infection showed a decrease in viral load after 12 months of follow-up in 30.2%, and in patients with cervical intraepithelial neoplasia grade I of older age — in 9.3%. Discussion. The management tactics for grade I cervical intraepithelial neoplasia associated with papillomavirus infection depends largely on the age of the patients and the degree of viral load. In young patients with immediate reproductive plans, a wait-and-see approach with dynamic observation for 12 months without drug therapy is possible in view of the probable spontaneous elimination of the human papillomavirus within a year. The results of the study suggest a higher rate of human papillomavirus elimination in cervical intraepithelial neoplasia grade I in women under 30 years of age. Conclusion. In cervical intraepithelial neoplasia of degree I in women under 40, infection with several types of the human papilloma virus (16, 18, 31, 51) with a clinically significant viral load (over 4.69±0.07 HPV per 100,000 cells) was found most frequently. In the group of the older reproductive age (40-49 years old), HPV HCC is less frequent in 6.90%. And the predominant role belongs to type 16, which was spread in 69.7% of cases.

Etiology of acute intestinal infections of viral character in the Altai territory

BACKGROUND: The Incidence of acute intestinal infections (AII) is generally maintained at a high level. Significant progress made in the field of laboratory diagnostics allowed us to proceed to a detailed study of the etiological structure of AII, and as a result, it was found that in recent years the role of pathogens of a viral nature has significantly increased, but a detailed study of the characteristics of these infections requires further research. AIMS: To study the structure and clinical and laboratory features of intestinal infections of viral etiology in adult hospitalized patients in the Altai Territory. MATERIAL AND METHODS: From 2017 to 2020, a single-stage study was conducted of 67 patients of an infectious hospital with AII of viral etiology, aged from 18 to 76 years, hospitalized in the infectious departments of the City Hospital No. 5, Barnaul. In order to determine the structure of AII, the polymerase chain reaction (PCR) method with hybridization-fluorescence detection AmpliSens AII screen-FL, bacteriological and serological methods were used. Taking into account the results of laboratory examination (molecular-genetic, bacteriological and serological), all patients were divided into 3 groups: group 1 ― 45 patients with viral monoinfection (67.2%), group 2 ― 9 patients with viral mixed infection (13.4%) and group 3 ― 13 patients with combined viral ― bacterial etiology (19.4%). Data processing and graphical representation were performed using computer programs Statistica 10.0 (Russian version) (Russia), Excel 2010 (Windows 10). RESULTS: As a result of the study, it was found that among the AII of viral etiology, 67.2% had monoviral AII and 32.8% had mixed AII-mixed viral (13.4%) and viral-bacterial etiology (19.4%). Among monoviral AII dominated norovirus infection (in 52,9%). Noted that the differences of clinical and laboratory parameters of the monoviral and mixed-viral infections have not been found, and there were no differences in groups of monoviral infections: rotavirus, adenovirus and norovirus, which is consistent with earlier data. As well as co-infection of salmonellosis, in combination with one and/or several viruses, more often occurred with febrile syndrome, watery diarrhea and abdominal pain. CONCLUSION: Thus, in adult patients in the conditions of an infectious hospital, mono-and mixed-virus infection (in combination with two or more viruses or bacteria) was detected in the structure of viral AII by PCR. There was no statistically significant difference in clinical and laboratory parameters in the study groups due to the insufficient sample size and the feasibility of conducting further studies using a similar protocol with a larger number of patients.