Molecular Genetic Techniques Research Articles

Molecular diagnostics of Cryptosporidium representatives in industrial pig industry in the Vologda region North-Western federal district of the Russian Federation

Cryptosporidiosis is a widespread parasitic disease of many species of domestic and wild animals, as well as humans, which is a significant problem in the field of medicine and veterinary medicine. Farm animals, in particular piglets, are most often susceptible to this disease, but the species composition of representatives of the genus Cryptosporidium in this species of animals in the Russian Federation has remained unknown to date. For the first time in the Russian Federation, in the conditions of the north-west, on the example of the Vologda region in pig farms with industrial technology for keeping piglets using the latest molecular genetic techniques, namely, using high-throughput sequencing of amplicon libraries of fragments of the 18S rRNA gene obtained as a result of nested PCR, we have established parasitism of C. scrofarum in all age groups of the animals examined. The total invasion of livestock in pig farms was 34% (51/150). Pine piglets were invaded by Cryptosporidium in 40% of cases (12/30), the intensity of Cryptosporidium infection was predominantly strong (+ + +), occurred in 20% (6/30) of cases. The mean (+ +) and weak (+) degree of animal invasion also occurred in 3% (10/30) of cases each. In the age group of weaning piglets at the age of 1-3 months, Cryptosporidium infection was 33.3% (10/30), a strong (+ + +) degree of oocyst production dominated, it was 26.7% (8/30) versus 10% (3/30) of the average (+ +) and 3.3% (1/30) weak (+). The most invasive oocysts are Cryptosporidium of fattening piglets at the age of 4-6 months, the extensinvasion of this group was 60% (18/30). The degree of oocyst release was predominantly moderate (+ +) – 26.7% (8/30) and strong – 23.3% (7/30). In 13.3% (4/30) of cases, there was a weak (+) degree of Cryptosporidium infection . Animals older than 6 months were infected with Cryptosporidium in 20% (6/30) of cases. They also recorded an average (+ +) – 13.3% (4/30) and a strong (+ + +) – 6.7% (2/30) degree of invasion, weak (+) invasion in this age group was not detected. Sows have also been invaded by Cryptosporidium. Their infection rate was 16.7% (5/30), and the degree of cryptosporidiosis was weak (+).

Molecular genetic testing in ovarian cancer

Ovarian cancer (OC) remains to be a leading cause of mortality among oncogynaecological patients. The low five-year survival rate of OC patients is associated with a lack of highly sensitive screening, early diagnostics and preventive methods, as well as high metastasis, recurrence and chemoresistance rates. Molecular genetic techniques for OC diagnosis based on standardized genetic panels can be used to detect a limited range of mutations in the BRCA1 and BRCA2 genes. However, the spectrum of genes potentially responsible for OC development is much wider. Recent data emphasize the importance of personalized approaches to account for ethno-population specifics in molecular genetic testing. This paper reviews recent data on the pathogenesis, molecular genetic diagnostic methods, and preventive strategies for OC.

Current <i>Streptococcus Pneumoniae</i> Serotypes in Sverdlovsk Region in 2020–2021

Relevance. Studying the spread of individual S. pneumoniae serotypes is of special epidemiologic and clinical importance. Sverdlovsk Region with the 20% proportion of community­-acquired pneumonia and its proportion among the causes of mortality of up to 70% is of special concern.Aims. Studying the features of the S. pneumoniae serotype landscape in individual socially significant population groups in Sverdlovsk Region in 2020 and 2021.Materials and methods. The work presents the results of testing nasopharyngeal smears collected in two observation groups, namely, that of 159 unvaccinated children aged 0 to 5 with acute respiratory infections (ARI), and that of 392 apparently healthy adults aged 19 to 27. We performed molecular genetic typing for identification of 20 leading S. рneumoniae serotypes/serogroups (1, 2, 3, 4, 5, 14, 16F, 19A, 19F, 23A, 23F, 6ABCD, 7AF, 9AV, 9LN, 11AD, 15AF, 18ABCF, 22AF, 12FAB/44/46, 33AF/37) using multiplex polymerase chain reaction (PCR).Results and discussion. In the individuals of the studied groups of observation, in 27.0% and 28.1% of the cases the study detected the S. pneumoniae in the biological material with circulation of 12 «vaccine» (19F – 2,6%, 6ABCD – 2,6%, 4 – 2,0%, 23F – 2,0%, 9AV – 1,3%, 9LN – 0,6%, 11AD – 1,3%, 18ABCF – 2,0%) and «nonvaccine» (15AF – 0.7%, 16F – 0.6%) serotypes. Nontypable strains with the highest proportion (77.3%) among the immunized adult population were predominant in the structure; in the unvaccinated children, this rate was 53.5%. Among the typable strains in children, 70.0% belonged to the vaccine ones, and most frequently the study identified 19F and 23F serotypes, while serotypes 3 and 12F/A/B/44/46 dominated among the adults.Conclusion. The study of unimmunized pediatric and immunized adult populations of Sverdlovsk Region confirmed a trend toward an increase in the circulation of nontypable S. pneumoniae strains in the Russian Federation, which sets the agenda to improve the monitoring system and optimize molecular genetic techniques for identification of the pathogens and upgrade the present specific and nonspecific preventive remedies, including the current antimicrobial ones and locally­-produced vaccine.

Molecular species identification of Rockskipper (Pisces: Blenniidae) from Porok Beach (Yogyakarta, Indonesia) based on 16S rRNA and COI genes

Rockskipper belongs to the species-rich family of Blenniidae (Combtooth blenny) and is known for its cryptic and species complexes presence. Identifying cryptic species based solely on morphological features is challenging due to their similar morphological characteristics. As a result, molecular genetic techniques based on two partial mtDNA genes, COI and 16S rRNA, were employed to identify fish accurately. This study aimed to evaluate the effectiveness of COI and 16S rRNA gene for the identification of Rockskipper fish and investigate the genetic relationship between species of Rockskipper from Porok Beach. The result revealed that the thirteen Rockskipper samples from Porok Beach that belong to six species (E. vermiculatus, E. striatus, I. lineatus, I. dussumieri, I. edentulus, and B. caudolineata) with more than 99% similarity. In contrast, the 16S analysis identified five species. A Bayesian phylogenetic tree demonstrated that six species of Rockskipper from Porok Beach are genetically distinct and separated into two clusters. We also found that two samples (RS-9 and RS-10) form a monophyletic group with B. caudolineata with maximum bootstrap (NJ and ML: 100%) and posterior probability (1.00). We hypothesized that B. caudolineata is a species complex with at least two lineages: one was genetically closer to RS-8, and another was more closely related to RS-9 and RS-10. Both the COI and 16S rRNA genes were found to be capable of delineating species and revealing genetic variation among Rockskipper samples in this study. However, our findings demonstrated that the COI gene is a more accurate and reliable marker for identifying Rockskipper species from Porok Beach, Gunungkidul, Yogyakarta, Indonesia.

The role of molecular-genetic techniques in BVDV eradication in Lower Austria.

A voluntary bovine viral diarrhoea virus (BVDV) control programme, which later became a compulsory eradication programme, based on the Swedish model was introduced in Lower Austria in 1997. The persistently infected animals were detected by Ag-ELISA and all samples were re-tested by the improved single-tube RT-PCR, employing panpestivirus primers targeting the 5'-UTR of the virus genome. In 2010, the BVDV eradication programme, which became compulsory from 2004, reached the final stage with only five remaining infected herds in which BVDV was difficult to eradicate. To resolve the problem in those herds, a molecular epidemiology approach was used. No differences in the spectrum of BVDV-1 subgenotypes at the beginning and at the final stage of eradication programme were found. The genetic study revealed the importance of human risk factor when finishing an eradication programme. Molecular epidemiology was also used to analyse BVDV isolates associated with re-introductions to BVDV-free herds.

The Emerging Roles of MicroRNAs as Biomarkers in Diabetic Nephropathy

Diabetic nephropathy (DN) is one of the most common complications of diabetes mellitus (DM), as well as the most common health issue among End Stage Renal Diseases (ESRD). Recent studies have shown that this concern will likely to become a global phenomenon in the future. Early diagnosis of DN is vital for its treatment. MicroRNAs (miRNAs or miRs) are the most promising for new biomarker candidates proposed for DN. MiRNAs, known as non protein-coding short-chain RNA sequences, have a regulatory role in many cellular events. Advances in molecular genetics and successful genomic techniques allow miRNAs to be used in the diagnosis of several diseases. As the DN diagnostic markers used today are insufficient for some cases, identifying new diagnostic markers is the basis of recent studies. Successful use of miRNAs in the areas of cancer, immunity and diabetes indicates that the DN perspective should be based on miRNA. In this review, miRNAs and their role in DN will be reviewed.

Bone and soft tissue tumors: clinicoradiologic-pathologicmolecular-genetic correlation of novel fusion spindled, targetable-ovoid, giant-cell-rich, and round cell sarcomas.

New entities in the classification of bone and soft tissue tumors have been identified by use of advanced molecular-genetic techniques,including next-generation sequencing. Clinicoradiologic and pathologic correlation supports diagnostic classification. Tumors from four morphologically grouped areas are selected to enhance diagnosis and awareness among the multidisciplinary team. These include select round cell tumors, spindle cell tumors, targetable tyrosine kinase/RAS::MAPK pathway-ovoid (epithelioid to spindled)tumors, and giant-cell-rich tumors of bone and soft tissue. Round cell tumors of bone and soft tissue include prototypical Ewing sarcoma, newer sarcomas with BCOR genetic alteration and CIC-rearranged, as well as updates onFUS/EWSR1::NFATc2, an EWSR1 non-ETS tumor that is solid with additional amplified hybridization signal pattern of EWSR1.This FUS/EWSR1::NFATc2 fusion hasnow been observed in seemingly benign to low-grade intraosseous vascular-rich and simple (unicameral) bone cyst tumors. Select spindle cell tumors of bone and soft tissue include rhabdomyosarcoma with FUS/EWSR1::TFCP2, an intraosseous high-grade spindle cell tumor without matrix. Targetable tyrosine-kinase or RAS::MAPK pathway-tumors of bone and soft tissue include NTRK, ALK, BRAF, RAF1, RET, FGFR1, ABL1, EGFR, PDGFB,and MET with variable ovoid myopericytic to spindled pleomorphic features and reproducible clinicopathologic and radiologic clues to their diagnosis. Giant-cell-rich tumors of bone, joint, and soft tissue are now respectivelycharacterized by H3F3A mutation, CSF1 rearrangement (targetable), and HMGA2::NCOR2 fusion. This article is an update for radiologists, oncologists, surgeons, and pathologists to recognize these novel ovoid, spindled, giant-cell-rich, and round cell tumors, for optimal diagnostic classification and multidisciplinary team patient care.

Use of Bacillus Strains Producing Lichenase and Laminarinase Enzymes as Fish Feed Additives

To obtain high efficiency from animals, it is necessary to maximize the ability to benefit from feed, as well as to protect animal health. One of the important applications in this direction is feed additives. Feed additives are plant and animal products and microorganisms that increase fish production and welfare, improve digestion and digestive system microflora and contribute to the preservation of nutrients and feed. Although plant and animal tissues are used in enzyme production, the use of microorganisms is in the first place. After the use of molecular genetic techniques, the genes responsible for enzyme production were cloned in microorganisms, making it possible to produce enzymes more purely and cheaply on an industrial scale.

A Review of Encephalitozoon cuniculi in Domestic Rabbits (Oryctolagus cuniculus)-Biology, Clinical Signs, Diagnostic Techniques, Treatment, and Prevention.

Encephalitozoon cuniculi is a eukaryote, unicellular, spore-forming, obligate intracellular microorganism of the phylum Microsporidia, with domestic rabbits as its main host. Another important species in which this pathogen has been identified are humans, the infection being therefore called a "zoonosis". The transmission takes place via the horizontal route or the vertical route, and cell-mediated immunity plays the biggest role in the infected hosts' protection. Encephalitozoonosis can manifest itself as an acute infection, with neurological signs, renal signs, and ocular lesions, or as a chronic or subclinical infection, which is usually the case for asymptomatic carriers. The diagnostic techniques usually carried out are histological examination, serological tests, and molecular genetic techniques. The treatment of encephalitozoonosis is usually symptomatic, with unrewarding results, and prevention methods include periodical serological screening, prophylactic administration of fenbendazole, and maintenance of a clean environment. The purpose of this article is to review the current data regarding the pathogenesis, host immunity, clinical signs, diagnostic methods, treatment, and prevention methods of encephalitozoonosis in the domestic rabbit, as well as to analyze the prevalence of this disease in different countries of the world.

A Blueprint for Genetic Determinism

A Blueprint for Genetic Determinism

Cytogenetic Analysis of Sporadic First-Trimester Miscarriage Specimens Using Karyotyping and QF-PCR: A Retrospective Romanian Cohort Study.

It is well known that first-trimester miscarriages are associated with chromosome abnormalities, with numerical chromosome abnormalities being the ones most commonly detected. Conventional karyotyping is still considered the gold standard in the analysis of products of conception, despite the extended use of molecular genetic techniques. However, conventional karyotyping is a laborious and time-consuming method, with a limited resolution of 5-10 Mb and hampered by maternal cell contamination and culture failure. The aim of our study was to assess the type and frequency of chromosomal abnormalities detected by conventional karyotyping in specimens of sporadic first-trimester miscarriages in a Romanian cohort, using QF-PCR to exclude maternal cell contamination. Long-term cultures were established and standard protocols were applied for cell harvesting, slide preparation, and GTG banding. All samples with 46,XX karyotype were tested for maternal cell contamination by QF-PCR, comparing multiple microsatellite markers in maternal blood with cell culture and tissue samples. Out of the initial 311 specimens collected from patients with sporadic first-trimester miscarriages, a total of 230 samples were successfully analyzed after the exclusion of 81 specimens based on unsuitable sampling, culture failure, or QF-PCR-proven maternal cell contamination. Chromosome abnormalities were detected in 135 cases (58.7%), with the most common type being single autosomal trisomy (71/135-52.6%), followed by monosomy (monosomy X being the only one detected, 24/135-17.8%), and polyploidy (23/135-17.0%). The subgroup analysis based on maternal age showed a statistically significant higher rate of single trisomy for women aged 35 years or older (40.3%) compared to the young maternal age group (26.1%) (p = 0.029). In conclusion, the combination of conventional karyotyping and QF-PCR can lead to an increased chromosome abnormality detection rate in first-trimester miscarriages. Our study provides reliable information for the genetic counseling of patients with first-trimester miscarriages, and further large-scale studies using different genetic techniques are required.

A versatile protoplast system and its application in Cannabis sativa L.

Cannabis sativa L. (cannabis) is one of the earliest domesticated crops cultivated for medicinal, food, industrial, and recreational use throughout the world. A growing number of jurisdictions have lifted the prohibition on medical and recreational cannabis with legislation that allows researchers to conduct important and urgently needed research, such as understanding the molecular basis of agronomically important traits. However, a dearth of suitable molecular genetic techniques for the analysis of cannabis species has hampered the elucidation of gene function. Here, we developed a robust protoplast transient transformation system by optimizing conditions of protoplast isolation and polyethylene glycol–mediated transformation using cannabis cotyledons. A protoplast isolation yield of up to 1.15 × 107 cells per gram of fresh material and 98.5% viability and a transformation efficiency of 75.4% were achieved. This optimized protocol for protoplast isolation and transformation was successfully applied to eight representative cannabis varieties, which highlight the versatility and robustness of this technique. Using this protocol, the nuclear localization of a basic helix–loop–helix transcription factor (CsMYC2) was visualized, and the transcriptional activity of CsMYC2 was assessed. These results demonstrate the effectiveness of our cannabis protoplast isolation and transient transformation method that should serve as a powerful molecular research tool in a wide range of molecular studies.

Genome-Wide Association Study Reveals Novel QTLs and Candidate Genes for Grain Number in Rice.

Grain number per panicle (GNPP), determined mainly by panicle branching, is vital for rice yield. The dissection of the genetic basis underlying GNPP could help to improve rice yield. However, genetic resources, including quantitative trait loci (QTL) or genes for breeders to enhance rice GNPP, are still limited. Here, we conducted the genome-wide association study (GWAS) on the GNPP, primary branch number (PBN), and secondary branch number (SBN) of 468 rice accessions. We detected a total of 18 QTLs, including six for GNPP, six for PBN, and six for SBN, in the whole panel and the indica and japonica subpanels of 468 accessions. More importantly, qPSG1 was a common QTL for GNPP, PBN, and SBN and was demonstrated by chromosome segment substitution lines (CSSLs). Considering gene annotation, expression, and haplotype analysis, seven novel and strong GNPP-related candidate genes were mined from qPSG1. Our results provide clues to elucidate the molecular regulatory network of GNPP. The identified QTLs and candidate genes will contribute to the improvement of GNPP and rice yield via molecular marker-assisted selection (MAS) breeding and genetic engineering techniques.

The Unusual Value of Long-Term Studies of Individuals: The Example of the Isle of Rum Red Deer Project

Long-term studies of individuals enable incisive investigations of questions across ecology and evolution. Here, we illustrate this claim by reference to our long-term study of red deer on the Isle of Rum, Scotland. This project has established many of the characteristics of social organization, selection, and population ecology typical of large, polygynous, seasonally breeding mammals, with wider implications for our understanding of sexual selection and the evolution of sex differences, as well as for their population dynamics and population management. As molecular genetic techniques have developed, the project has pivoted to investigate evolutionary genetic questions, also breaking new ground in this field. With ongoing advances in genomics and statistical approaches and the development of increasingly sophisticated ways to assay new phenotypic traits, the questions that long-term studies such as the red deer study can answer become both broader and ever more sophisticated. They also offer powerful means of understanding the effects of ongoing climate change on wild populations.

Impaired trafficking and instability of mutant kidney anion exchanger 1 proteins associated with autosomal recessive distal renal tubular acidosis

BackgroundMutations in solute carrier family 4 member 1 (SLC4A1) encoding anion exchanger 1 (AE1) are the most common cause of autosomal recessive distal renal tubular acidosis (AR dRTA) in Southeast Asians. To explain the molecular mechanism of this disease with hematological abnormalities in an affected family, we conducted a genetic analysis of SLC4A1 and studied wild-type and mutant AE1 proteins expressed in human embryonic kidney 293T (HEK293T) cells.MethodsSLC4A1 mutations in the patient and family members were analyzed by molecular genetic techniques. Protein structure modeling was initially conducted to evaluate the effects of mutations on the three-dimensional structure of the AE1 protein. The mutant kidney anion exchanger 1 (kAE1) plasmid construct was created to study protein expression, localization, and stability in HEK293T cells.ResultsWe discovered that the patient who had AR dRTA coexisting with mild hemolytic anemia carried a novel compound heterozygous SLC4A1 mutations containing c.1199_1225del (p.Ala400_Ala408del), resulting in Southeast Asian ovalocytosis (SAO), and c.1331C > A (p.Thr444Asn). Homologous modeling and in silico mutagenesis indicated that these two mutations affected the protein structure in the transmembrane regions of kAE1. We found the wild-type and mutant kAE1 T444N to be localized at the cell surface, whereas the mutants kAE1 SAO and SAO/T444N were intracellularly retained. The half-life of the kAE1 SAO, T444N, and SAO/T444N mutants was shorter than that of the wild-type protein.ConclusionThese results suggest impaired trafficking and instability of kAE1 SAO/T444N as the likely underlying molecular mechanism explaining the pathogenesis of the novel SLC4A1 compound heterozygous mutation identified in this patient.

Recent advances in gene therapy: genetic bullets to the root of the problem.

Genetics and molecular genetic techniques have changed many perspectives and paradigms in medicine. Using genetic methods, many diseases have been cured or alleviated. Gene therapy, in its simplest definition, is application of genetic materials and related techniques to treat various human diseases. Evaluation of the trends in the field of medicine and therapeutics clarifies that gene therapy has attracted a lot of attention due to its powerful potential to treat a number of diseases. There are various genetic materials that can be used in gene therapy such as DNA, single- and double-stranded RNA, siRNA and shRNA. The main gene editing techniques used for in vitro and in vivo gene modification are ZNF, TALEN and CRISPR-Cas9. The latter has increased hopes for more precise and efficient gene targeting as it requires two separate recognition sites which makes it more specific and can also cause rapid and sufficient cleavage within the target sequence. There must be carriers for delivering genes to the target tissue. The most commonly used carriers for this purpose are viral vectors such as adenoviruses, adeno-associated viruses and lentiviruses. Non-viral vectors consist of bacterial vectors, liposomes, dendrimers and nanoparticles.

Quantitative Experimental Embryology: A Modern Classical Approach.

Experimental Embryology is often referred to as a classical approach of developmental biology that has been to some extent replaced by the introduction of molecular biology and genetic techniques to the field. Inspired by the combination of this approach with advanced techniques to uncover core principles of neural crest development by the laboratory of Roberto Mayor, we review key quantitative examples of experimental embryology from recent work in a broad range of developmental biology questions. We propose that quantitative experimental embryology offers essential ways to explore the reaction of cells and tissues to targeted cell addition, removal, and confinement. In doing so, it is an essential methodology to uncover principles of development that remain elusive such as pattern regulation, scaling, and self-organisation.

Taxonomic Reclassification of the Fungal Pathogen Causing Dry Berry Disease of Caneberries into the Division Ascomycota as Monilinia rubi.

As molecular genetic techniques improve and sequence data becomes available for more fungal species, taxonomic classifications historically based upon growth morphology alone are being revisited and occasionally reclassified. Herein, we present such an instance for the fungal pathogen that causes dry berry disease of caneberries. The organism was previously described as the basidiomycete fungus Rhizoctonia rubi based upon the pathogen's production of Rhizoctonia-like angular branching hyphae. Utilizing molecular genetic techniques unavailable when the pathogen was first characterized in 1959, three housekeeping gene regions (ITS, β-tubulin, and G3PDH) were sequenced across 13 contemporary dry berry isolates, as well as the original 1959 R. rubi type strain, CBS382.59. The resulting neighbor-joining, maximum likelihood, and Bayesian phylogenies for single and multilocus sequences provide strong evidence that the dry berry pathogen was misclassified. This data, in addition to revisiting invivo macroscopic and microscopic growth morphology, again comparing contemporary dry berry isolates to the CBS382.59 type strain, suggests that the causal organism is a new species within the genus Monilinia that we propose be classified as Monilinia rubi. A transition from designation as a basidiomycete fungus to an ascomycete fungus could have implications on chemical management decisions, as well as the assumptions made about cell structure and the pathogen's putative life cycle.

Genetic risk of osteoarthritis operates during human skeletogenesis.

Osteoarthritis (OA) is a polygenic disease of older people resulting in the breakdown of cartilage within articular joints. Although it is a leading cause of disability, there are no disease-modifying therapies. Evidence is emerging to support the origins of OA in skeletogenesis. Whereas methylation quantitative trait loci (mQTLs) co-localizing with OA genome-wide association study signals have been identified in aged human cartilage and used to identify effector genes and variants, such analyses have never been conducted during human development. Here, for the first time, we have investigated the developmental origins of OA genetic risk at seven well-characterized OA risk loci, comprising 39 OA-mQTL CpGs, in human fetal limb (FL) and cartilage (FC) tissues using a range of molecular genetic techniques. We identified significant OA-mQTLs at 14 and 29 CpGs in FL and FC tissues, respectively, and compared our results with aged cartilage samples (AC). Differential methylation was observed at 26 sites between FC and AC, with the majority becoming actively hypermethylated in old age. Notably, 6/9 OA effector genes showed allelic expression imbalances during fetal development. Finally, we conducted ATAC-sequencing in cartilage from the developing and aged hip and knee to identify accessible chromatin regions and found enrichment for transcription factor binding motifs including SOX9 and FOS/JUN. For the first time, we have demonstrated the activity of OA-mQTLs and expression imbalance of OA effector genes during human skeletogenesis. We show striking differences in the spatiotemporal function of these loci, contributing to our understanding of OA aetiology, with implications for the timing and strategy of pharmacological interventions.

RESULTS OF MONITORING THE INCIDENCE OF NON-POLIO ENTEROVIRUS INFECTION IN THE PRE-PANDEMIC AND PANDEMIC PERIODS

The results of monitoring the incidence of non-polio enterovirus infection (EVI) in the pre-pandemic and pandemic periods in the Khanty-Mansiysk Autonomous Okrug (2006-2021) are presented. It was found that the predominant share in the total incidence of EVI was urban population with a predominance of children under 6 years of age and school age. In the period 2020-2021. The low incidence of EVI was due to ongoing anti-epidemic measures against COVID-19 and was accompanied by the spread of enterovirus serotype CVA6, which is able to effectively implement the aspiration mechanism of transmission of the infectious agent and cause respiratory and exanthemic diseases. The spread of CVA6 was influenced by the growth of migration activity of the population, due to the partial lifting of restrictions and an increase in the non-immune stratum of the population due to a decrease in the incidence of EVI in 2020. The results of the study will contribute to the adoption of measures leading to a decrease in the incidence of EVI in the Khanty-Mansiysk Autonomous Okrug, rational planning of preventive measures, by improving the information and diagnostic subsystems of epidemiological surveillance. To solve these tasks, it is necessary to further study the spectrum of enteroviruses circulating on the territory of the Khanty-Mansi Autonomous Okrug, the frequency of their occurrence in subclinical forms of EVI and as etiological agents of enterovirus meningitis, the development of molecular genetic typing techniques using mass parallel high-performance sequencing technology, the study of molecular genetic characteristics. All this will reduce the risks of EVI morbidity in the post-pandemic period and ensure the sanitary and epidemiological well-being of the population and prevent the spread of EVI in the Khanty-Mansi Autonomous Okrug.