Problemy Osobo Opasnykh Infektsii最新文献

{"title":"Crimean-Congo Hemorrhagic Fever: Epidemiological and Epizootiological Situation in the Russian Federation in 2022, Incidence Forecast for 2023","authors":"A. Volynkina, N. Tkachenko, O. Maletskaya, O. N. Skudareva, I. V. Tishchenko, A. A. Zhirova, Y. Lisitskaya, L. Shaposhnikova, D. V. Rostovtseva, E. Manin, D. Prislegina, V. Petrovskaya, E. V. Yatsmenko, A. Kulichenko","doi":"10.21055/0370-1069-2023-2-6-12","DOIUrl":"https://doi.org/10.21055/0370-1069-2023-2-6-12","url":null,"abstract":"The review presents an analysis of the epidemiological and epizootiological situation on Crimean-Congo hemorrhagic fever (CCHF) in the Russian Federation in 2022. The incidence rate of CCHF registered in 2022 (59 cases) was 1.2 times higher as compared to 2021, however, below the long-term average annual values. The mortality rate was 10.2 %, which exceeds the indicators of long-term observations (in 2012–2021 – 3.2 %). Following epizootiological survey of stationary observation points, it was found that the number of Hyalomma marginatum imago in 2022, in general, corresponded to the average long-term indicators. CCHF virus isolates circulating in Russia in 2017–2022 belonged to the genetic lines “Europe-1” (V), “Europe-2” (VI), and “Europe-3” (VII). The ratio of CCHF virus genovariants in the population on the territory of the Russian Federation in 2017–2022 didn’t change. Based on the analysis of naturalclimatic factors, the forecast for the incidence of CCHF in the Russian Federation for 2023 has been made.","PeriodicalId":52264,"journal":{"name":"Problemy Osobo Opasnykh Infektsii","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45074473","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Paleogenomics of the Plague Agent and Prospects for Paleogenomic Studies in Russia","authors":"G. A. Eroshenko, E. Batieva, V. Kutyrev","doi":"10.21055/0370-1069-2023-2-13-28","DOIUrl":"https://doi.org/10.21055/0370-1069-2023-2-13-28","url":null,"abstract":"The review contains information on paleogenomic studies of the plague pathogen, Yersinia pestis, covering the prehistoric epoch, the periods of the first and second plague pandemics, epidemics and outbreaks of plague of the late XIX–XX centuries. We have summarized the data on the reconstruction of ancient Y. pestis genomes of the Late Neolithic, Bronze and Iron Ages, the Justinian Plague epidemic of the first plague pandemic, the Black Death epidemic and the subsequent epidemics of the second plague pandemic of the XIV–XVIII centuries, as well as on tracing the pathways of plague propagation waves in Eurasia and the course of plague agent evolution with the formation of a vector-borne transmission route with the help of arthropods. We present the results of our own research of Y. pestis genomes from the key sites of formation of etiological agents of the first and second plague pandemics in the Tien Shan Mountains, historical outbreaks in the Northern and North-Western Caspian sea region and other regions of Russia and adjoining countries in the late XIX–XX centuries. The paper discusses the areas of the Caucasus, Crimea, Northern Caspian, Siberia, and Tien Shan in the territory of Russia and neighboring states that are promising for national paleogenomic studies of plague.","PeriodicalId":52264,"journal":{"name":"Problemy Osobo Opasnykh Infektsii","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43323436","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"New Method of Vaccination Against Anthrax","authors":"A. Shevtsov, S. A. Permyakov, N. G. Khapaev, R. S. Ziganshin","doi":"10.21055/0370-1069-2023-1-157-163","DOIUrl":"https://doi.org/10.21055/0370-1069-2023-1-157-163","url":null,"abstract":"The aim of this work was to develop an oral method of vaccination against anthrax, since wide vaccination campaigns using this method require no special conditions, equipment, and instruments. Also, minimum number of medical personnel is sufficient to ensure control over the intake of the drug. Materials and methods. Domestic raw materials, consumables and reagents that passed the incoming inspection and met the requirements of State and industry standards, technical specifications, and the articles of the Pharmacopoeia were used for the work. Batches of anthrax vaccines based on Bacillus anthracis strain STI-1, manufactured at the Research Center (Kirov) of the “48th Central Research Institute” of the Ministry of Defense of Russia were put to the test. Results and discussion. The immunization schemes have been tested on animals, taking into account the features of the antigen. One of the crucial factors that determine the effectiveness of oral vaccination is the correct choice of the type and form of the oral anthrax vaccine administered enterally. It has been shown that in contrast to oral vaccines, the vaccine strain in enteral vaccines must be protected from the harmful effects of stomach contents. Thus, enteric-coated capsules coated with a shell that is resistant to the action of stomach acid were used for the study. The presented experimental data indicate that a single administration of the capsules with oral anthrax vaccine STI protects at least 70 % of laboratory animals from a highly virulent strain of the anthrax microbe and confirm the safety and non-reactogenicity of the drug. The developed laboratory technology makes it possible to obtain a finished product containing one inoculation dose of an oral vaccine for laboratory animals. So, a new method of vaccination has been designed. It is necessary to conduct preclinical and clinical trials to promptly introduce the oral administration of anthrax vaccine into medical practice as the simplest method for mass vaccination of humans.","PeriodicalId":52264,"journal":{"name":"Problemy Osobo Opasnykh Infektsii","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47569403","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Typing of Uncultured Isolates of Coxiella burnetii and Coxiella-Like Microorganisms Associated with Ticks Using 16S rRNA Gene Nucleotide Sequence Analysis","authors":"Y. Panferova, N. Tokarevich, O. Blinova, A. A. Nafeev, E. I. Sibaeva","doi":"10.21055/0370-1069-2023-1-142-147","DOIUrl":"https://doi.org/10.21055/0370-1069-2023-1-142-147","url":null,"abstract":"The causative agent of Q fever, the intracellular pathogen Coxiella burnetii, is found almost worldwide; many types of blood-sucking ticks that are dangerous to animals and humans are involved in the circulation of the pathogen. Using molecular-genetic methods, closely related species of microorganisms of the genus Coxiella sp. have been discovered, some of which are endo-symbionts of ticks, and some can survive in the human body, causing an infectious process. The existence of species whose genes are similar in nucleotide sequence to those of C. burnetii makes it difficult to diagnose the pathogen in arthropod vectors. The aim of this work was to consider the use of PCR and sequencing of an extended 16S rRNA gene fragment for molecular diagnostics and differentiation of C. burnetii from Coxiella-like microorganisms. Materials and methods. Individual samples of blood-sucking ticks were examined to detect bacteria of the genus Coxiella sp. applying standard PCR. For positive samples, an extended fragment of the 16S rRNA gene was obtained and examined by sequencing and multiple alignment with homologous sequences. Results and discussion. Of the 96 examined ticks collected in the Ulyanovsk Region, one was positive for the presence of C. burnetii DNA and one – for the presence of Coxiella sp. The greatest similarity for the C. burnetii isolate was noted in comparison with Western European strains, for the Coxiella-like microorganism - with closely related bacteria from ticks of the same species. Unique polymorphisms for the detected microorganisms were identified. It has been established that genus-specific primers to the 16S rRNA gene fragment are able to amplify not only bacteria of the genus Coxiella sp., but also genetically distant species. Analysis of the sequence of the extended 16S rRNA gene fragment makes it possible to differentiate C. burnetii from Coxiella-like microorganisms; some gene polymorphisms appear to have arisen through microevolution in different geographic regions. In the European part of the Russian Federation, Coxiella-like bacteria have been uncovered for the first time.","PeriodicalId":52264,"journal":{"name":"Problemy Osobo Opasnykh Infektsii","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46224034","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Results of a Joint Epizootiological Survey of Transboundary Natural Plague Foci of the Russian Federation and Potentially Focal Territories of the Republic of Kazakhstan in 2019–2022","authors":"A. M. Senichkina, I. Sharova, S. V. Magerramov, K. S. Zakharov, A. A. Kuznetsov, V. N. Chekashov, A. Porshakov, M. G. Korneev, M. V. Proskuryakova, Yu.V. Kislitsyn, A. Belyaev, I. B. Kim, L. Nurmagambetova, I. G. Kozulina, A. A. Bashmakov, A. A. Bashmakova, A. A. Kovalevskaya, R. Adilov, M. P. Grigor’ev, E. Kuklev, A. K. Grazhdanov","doi":"10.21055/0370-1069-2023-1-148-156","DOIUrl":"https://doi.org/10.21055/0370-1069-2023-1-148-156","url":null,"abstract":"Consolidation of the efforts in implementation of epidemiological surveillance and control over plague and other dangerous natural-focal infections is an essential aspect in ensuring epidemiological well-being as regards particularly dangerous infectious diseases in the territory of natural plague foci and potentially focal territories located within the borders of the Russian Federation and the Republic of Kazakhstan. The aim of the work was to carry out a joint epizootiological survey of the transboundary territories of the Volga-Ural sandy natural plague focus and the territory of the East Kazakhstan region of the Republic of Kazakhstan (RK) potentially focal for plague over the period of 2019–2022. Materials and methods. Samples of field materials, collected during the epizootiological survey of the territory of Kazakhstan, were studied using bacteriological, molecular-genetic, and immune-serological methods. Results and discussion. We have obtained the current evidence on the spatial-biocenotic structure, the circulation of pathogens of dangerous natural-focal infectious diseases in the transboundary territories of Eastern and Western Kazakhstan. It has been established that the conditions that contribute to the possibility of human infection with plague and other dangerous infectious diseases in case of the aggravation of epizootic situation in the foci or importation of the pathogens into the territory are in place.","PeriodicalId":52264,"journal":{"name":"Problemy Osobo Opasnykh Infektsii","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45864914","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Effect of COVID‑19 on the Incidence of Human Vector-Borne Infections Transmitted by Ixodid Ticks (Illustrated by the Example of Krasnoyarsk Territory)","authors":"A. Nikitin, O. V. Sorokina, E. I. Andaev, N. Y. Kharlamp’eva, S. Balakhonov","doi":"10.21055/0370-1069-2023-1-120-125","DOIUrl":"https://doi.org/10.21055/0370-1069-2023-1-120-125","url":null,"abstract":"In 2022, there was an increase in the incidence of tick-borne encephalitis (TBE) and ixodid tick-borne borreliosis (ITBB) in the subjects of the Russian Federation, federal districts and the country as a whole. At the same time, among people affected by ixodid ticks, the proportion of clinical forms of the diseases has increased. The aim of this study is to analyze the possible causes of the simultaneous increase in the incidence of vector-borne infections and the proportion of clinical forms among the people bitten by the ticks, using the materials on Krasnoyarsk Territory as an example. Materials and methods. We analyzed the incidence of TBE and ITBB among the people who have or haven’t had COVID‑19, the number of people affected by tick bites in 2021–2022 in Krasnoyarsk Territory. Statistical analysis is performed by standard methods of variational statistics using Excel. Results and discussion. It is shown that in the Krasnoyarsk Territory, with a decrease in the number of individuals affected by tick bites in 2022 as compared to 2021 (12216 and 13214, respectively), there was an increase in the number of cases of TBE (from 124 to 250) and ITBB (from 115 to 224), as well as the proportion of people who had clinical forms of the infections after tick bites. Among three possible explanations of the observed pattern, a decrease in the immune status of Krasnoyarsk residents who had had COVID‑19 before the contact with ticks is considered as the most likely one. Using statistical methods it was substantiated that among people with TBE and ITBB, the proportion of individuals who have had COVID‑19 is significantly higher than expected, based on the actual incidence of the Krasnoyarsk Territory population with a new coronavirus infection. Thus, COVID‑19 affects the incidence of vector-borne infections indirectly, through a change in the number of people contacts with natural stations, and directly, by changing the immune status.","PeriodicalId":52264,"journal":{"name":"Problemy Osobo Opasnykh Infektsii","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47390695","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Intraspecific Differentiation of Francisella tularensis Strains Using Multilocus Real-Time Polymerase Chain Reaction","authors":"N. A. Osina, D. A. Sitmbetov, E. G. Bulgakova, S. S. Chekmareva, E. V. Sazanova, A. M. Senichkina, O. Y. Lyashova, A. Osin, S. A. Shcherbakova","doi":"10.21055/0370-1069-2023-1-132-141","DOIUrl":"https://doi.org/10.21055/0370-1069-2023-1-132-141","url":null,"abstract":"The aim of the study was to develop a method for intraspecific differentiation of the tularemia microbe: subspecies tularensis (subpopulations AI and AII), holarctica (biovars japonica, EryS/R), mediasiatica, and novicida using multilocus real-time PCR. Materials and methods. We used 48 strains of F. tularensis of various subspecies, biovars, and subpopulations. Intraspecific appurtenance of the strains was carried out on the basis of the analysis of the RD-1 region variability applying PCR, the sdhA gene by Sanger fragment sequencing and by the disk diffusion method using disks with erythromycin. The selection of primers and probes was performed using the software available at www.genscript.com and GeneRunner 6.5.52. Sequence homology was assessed using the BLAST algorithm and the GenBank NCBI database. Results and discussion. New data on the structure and occurrence of the differentiation regions RD-8, RD-12, RD-28 of FTT1122c gene and its homologous sequences in strains of tularemia microbe of various subspecies have been obtained. Novel RDhm 346 bp in size, characteristic of strains of the subsp. mediasiatica, holarctica, which is deleted in subsp. tularensis and absent in subsp. novicida has been detected. Based on the detection of the FTT1670, FTT1122с, FTT1067, FTW_2084 loci, a multilocus real-time PCR has been developed – “F. tularensis 4c”, providing for identification of all subspecies of the tularemia microbe, separately for the biovar japonica of the Holarctic subspecies and subpopulations AI, AII of the subspecies tularensis. The PCR specificity was confirmed in the study of strains of tularemia microbe from the fund of the “State Collection of Pathogenic Bacteria” at the premises of the Russian Reserarch Anti-Plague Institute “Microbe”. The results obtained expand the concept of intraspecific genetic heterogeneity of tularemia microbe and possibilities of identifying the causative agent of tularemia using molecular-genetic methods. They are important for understanding the processes of adaptation of the pathogen to circulation in the host organism and environmental objects, the course of evolution and formation of new species of Francisella.","PeriodicalId":52264,"journal":{"name":"Problemy Osobo Opasnykh Infektsii","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44649593","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of an Integrated System for Molecular-Genetic Identification of Yersinia pestis Strains","authors":"K. A. Nikiforov, E. G. Oglodin, M. A. Makashova, A. N. Balykova, D. V. Utkin, L. M. Kukleva, G. A. Eroshenko, V. Kutyrev","doi":"10.21055/0370-1069-2023-1-126-131","DOIUrl":"https://doi.org/10.21055/0370-1069-2023-1-126-131","url":null,"abstract":"The paper describes a developed comprehensive system for molecular-genetic identification of Yersinia pestis strains according to their appurtenance to certain subspecies, biovars, phylo-geographic populations, using realtime PCR (RT-PCR), allele-specific RT-PCR, and multiplex PCR with hybridization fluorescent registration of results on a solid substrate. Application of this system makes it possible to establish the appurtenance of Y. pestis strains to the following phylogenetic branches: 0.ANT1, 0.ANT2, 0.ANT3, 0.ANT5, 3.ANT, 4.ANT of antique biovar of the main subspecies; 2.MED0, 2.MED1, 2.MED2, 2.MED3, 2.MED4 of medieval biovar of the main subspecies; 1.IN1, 1.IN2, 1.IN3 of intermedium biovar of the main subspecies; 1.ORI1, 1.ORI2, 1.ORI3 of oriental biovar of the main subspecies; 0.PE3 (angolica subspecies), 0.PE7 (tibetica subspecies) and 0.PE10 (qinghaica subspecies). The first stage of the studies within the frames of the developed system is indication of plague agent using registered diagnostic drugs. The second stage is the determination of belonging to individual subspecies through RT-PCR or by the method of multiplex PCR system with hybridization-fluorescent registration of results on a solid substrate, which also allows for establishing to which biovars of the main subspecies and the main phylogenetic lines of the ancient biovar the strains belong. The third stage is the identification of strain appurtenance to phylogenetic branches by the AS-RT-PCR method. The designed complex system for molecular-genetic identification of Y. pestis strains can be applied at the regional and federal levels of the laboratory network of the Russian Federation for diagnostics of infectious diseases. Its use will considerably facilitate and increase the efficiency of intraspecific differentiation of Y. pestis strains within the framework of the epidemiological investigation of outbreaks or importation of strains of plague pathogen into the territory of the Russian Federation or during the certification of strains in collection activities.","PeriodicalId":52264,"journal":{"name":"Problemy Osobo Opasnykh Infektsii","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41807152","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Population Genomics of SARS-CoV-2 in the Constituent Entities of Siberian and Far Eastern Federal Districts","authors":"L. V. Mironova, A. N. Bondaryuk, E. A. Sidorova, N. O. Bochalgin, I. S. Fedotova, Y. Bukin, A. S. Ponomareva, E. I. Andaev, S. Balakhonov","doi":"10.21055/0370-1069-2023-1-111-119","DOIUrl":"https://doi.org/10.21055/0370-1069-2023-1-111-119","url":null,"abstract":"The aim of the study was to analyze the genetic structure of populations and the patterns of evolutionary variability of the novel coronavirus infection in the Siberian and Far Eastern Federal Districts. Materials and methods. 1033 SARS-CoV-2 genomes from samples from individuals diagnosed with COVID-19 from eight entities of the Siberia and Far East between December 2020 and November 2021 were assessed. Sequencing was performed on the MinION Oxford Nanopore platform using the ARTIC v.3 protocol. The degree of SARS-CoV-2 genetic isolation was estimated applying the Fst criterion. Phylogenetic analysis was carried out using maximum likelihood method and Bayesian phylogenetic inference. A nonparametric Bayesian Skyline Plot (BSP) model was used to reconstruct population dynamics. Results and discussion. The original SARS-CoV-2 variant (B.1) was identified in 100 % of the cases at the initial stages. The Alpha variant was detected in March-June, 2021; Beta – in single samples in March-May, 2021. Delta was first identified in April, 2021. The maximum degree of SARS-CoV-2 genetic isolation (Fst=0.18) was established for the most remote territories (Altai Territory ↔ Republic of Buryatia and Altai Territory ↔ Irkutsk Region). A relatively free circulation of the virus was detected between Irkutsk Region, Republic of Buryatia and Krasnoyarsk Territory. According to the results of population genetic tests, a sharp increase in the effective virus population size was the determining mechanism of SARS-CoV-2 genetic diversity formation. Reconstruction of population dynamics in BEAST (BSP model) has revealed the consistency of trends in the genetic diversity of the virus and the number of active cases. Two subclusters have been identified in the Delta cluster, consisting predominantly of samples isolated in the Irkutsk Region and Krasnoyarsk Territory. Change in the dominant variant of SARS-CoV-2 has been traced in dynamics. Molecular-epidemiological data point to the multiple pathways of spatial expansion of different SARS-CoV-2 genotypes into the constituent entities with generation of individual monophyletic clusters and further intra- and extraterritorial spread of the decedents.","PeriodicalId":52264,"journal":{"name":"Problemy Osobo Opasnykh Infektsii","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48971859","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Obtaining and Characterization of Hybridomas Producing Monoclonal Antibodies against Coronavirus SARS-CоV-2","authors":"G. V. Kuklina, S. S. Ipatov, A. S. Gorshkov, D. Pechenkin, A. V. Eremkin, A. V. Kuznetsovsky, A. Tumanov, I. V. Darmov","doi":"10.21055/0370-1069-2023-1-105-110","DOIUrl":"https://doi.org/10.21055/0370-1069-2023-1-105-110","url":null,"abstract":"The aim of the work was to obtain and characterize hybridomas producing monoclonal antibodies to antigens of coronavirus SARS‑CoV‑2, promising for the construction of diagnostic immunochemical tests. Materials and methods. Recombinant nucleocapsid and receptor binding fragment of spike protein of SARS‑CoV‑2 were used for immunization of BALB/c mice. Antigens were absorbed on aluminium hydroxide gel and injected subcutaneously to BALB/c mice at a 7-day-interval. Immune splenocytes and myeloma cells SP2/0-Ag14 were fused by polyethylene glycol 1450. Cell cultures producing specific antibodies against nucleocapsid and receptor binding fragment were selected applying indirect ELISA in 96-well plates sensitized by desired antigens. Clones of hybridomas were obtained using the method of limiting dilutions. Production properties were studied through in vitro cultivation in 24-well culture plates. Immune-ascitic fluids were collected during the cultivation of hybrid cells in peritoneal cavities of BALB/c mice. Monoclonal antibodies were purified by affinity chromatography on protein A sepharose sorbent, conjugated with horseradish peroxidase, and tested for the possibility to be used in sandwich ELISA for detection of inactivated SARS‑CoV‑2 coronavirus strain “Isolate B”. Results and discussion. As a result of hybridization and selection of clones, hybridomas producing monoclonal antibodies to nucleocapsid and receptor binding fragment of SARS‑CoV‑2 have been obtained. During the in vitro and in vivo cultivation the clones maintained the consistent proliferative and antibody producing activity. The application of monoclonal antibody 415D12 as a capture one and 411D12 antibody conjugated with horseradish peroxidase as a detector antibody in ELISA allows for identifying SARS‑CoV‑2 coronavirus at a minimum concentration of 1·103 PFU per ml.","PeriodicalId":52264,"journal":{"name":"Problemy Osobo Opasnykh Infektsii","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48409372","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}