[Development and assessment of a reagent kit for RNA detection of CrimeanCongo hemorrhagic fever virus with using reverse transcription loop-mediated isothermal amplification method.]

This study presents the results of laboratory trials of the reagent kit for the rapid detection of RNA of the Crimean-Congo hemorrhagic fever virus (CCHFV) using loop-mediated isothermal amplification with reverse transcription (RT-LAMP). The developed RT-LAMP reagent kit was used to detect the CCHFV and showed a sensitivity of 103 GE/ml of viral RNA, which is sufficient for detection of the CCHFV in the early stage of human infections. The kit showed high specificity and no cross-reactivity with viral panel from the State collection of viruses of the FBRI SRC VB «Vector¼ (arboviruses and hemorrhagic fever viruses). Laboratory trials of the RT-LAMP kit are showed a high analytical and diagnostic sensitivity and specificity for RNA detection of the CCHFV and high speed of the analysis (60-70 min with sample preparation) compared to real-time PCR. Approbation of the kit field version has showed the possibility of setting the RT-LAMP reaction and viral RNA detection without the using of analytical equipments.

[EPIZOOTOLOGIC MONITORING OF NATURAL-FOCAL INFECTIONS IN THE SOUTH OF RUSSIA IN 2015].

AIM: Analyze epizootologic manifestations of natural-focal infections (NFI) in the south of Russia in 2015. MATERIALS AND METHODS: Reports provided by administrations of Federal Service of Surveillance for Protection of Consumers Rights and Human Welfare, Centers of Hygiene and Epidemiology, Research Institutes for Plague Control and Station for Plague Control were used. Data processing was carried out using Excel program. RESULTS: A total of 37-738 samples of field material were studied, most of those - for tularemia (8268), Crimea hemorrhagic fever (6894) and West Nile fever (4875). Causative agents of 15 nosologic forms of NFI were detected during epizootologic examination of the territory of south of Russia, predominating amount of positive samples was obtained during study of material for tick-borne borreliosis (612), Crimea hemorrhagic fever (279) and tularemia (237). CONCLUSION: Analysis of epizootologic situation on natural- focal infections in the south of Russia gives evidence on the increase of activity of natural foci of bacterial (tularemia, tick-borne borreliosis), rickettsia (tick-borne spotted fevers) and viral infec- tions (Crimea hemorrhagic fever, West Nile fever, tick-borne viral encephalitis), and thus problems of prophylaxis of natural-focal infectious diseases and constant monitoring of natural foci in su- bjects of Southern, North-Caucasian and Crimea federal districts are of particular importance.

[DETERMINATION OF PHYLOGENETIC RELATIONSHIP OF YERSINIA PESTIS STRAINS FROM NATURAL PLAGUE FOCI OF THE CAUCASUS BY MULTI-LO- CUS VNTR-ANALYSIS].

AIM: Determination of the degree of phylogenetic relationship of Yersinia pestis strains iso- lated from the territories of natural foci of plague from the Caucasus using VNTR-typing by 25 loci (MLVA25). MATERIALS AND METHODS: 26 strains of Y pestis from Russian natural foci of the Caucasus were used in the study. 25 loci of tandem repeats in Y pestis genome by Le Fleche scheme were used for execution of multi-locus VNTR-analysis. Deciphering of nucleotide sequences was carried out in automatic sequencer ABI 3130 Genetic Analyser. Analysis of confinement of clusters to certain territories, objects and time of isolation of strains was carried out. using Arc GIS 10.1 program. RESULTS: Groups of MLVA25-types of various levels of discrimination were formed: clusters, groups and subgroups. Clusters were formed by strains ofvarious taxonomic membership: main and subspecies of Y pestis. Subgroups reflect membership of strains in certain foci, and MLVA25-types - the degree of genetic relationship. CONCLUSION: Genetic <> of plague causative agents obtained using MLVA25-types circulating in various natural-focal territories allow to solve problems ofboth theoretical and practical character: from interpretation of microevolution processes to the search of the source of infection and ways of its spread during possible epidemic complications.

[RETROSPECTIVE ANALYSIS OF BIOLOGICAL AND MOLECULAR-GENETIC PROPERTIES OF STRAINS - CAUSATIVE AGENTS OF CHOLERA - ISOLATED IN UKRAINE IN 1994 - 2011].

AIM: Retrospective analysis of biological and molecular-genetic properties of strains - cau- sative agents of cholera - isolated in the period of epidemics in Ukraine in 1994 - 2011. MATERIALS AND METHODS: Phenotypic and molecular-genetic properties of 5 strains of cholera vibrios, biovar El Tor isolated from cholera patients and 4 strains from the environmental samples were studied using traditional bacteriological and genetic methods. Detection of DNA for toxigenicity genes and genes characteristic for El Tor and classic biovar were carried out by PCR method using rea- gent kits <> and <> an experimental test system). Sequencing of genomes of 4 strains of causative agents of cholera was carried out in genetic analyzer Ion Torrent Personal Genome Machine. RESULTS: Strains of cholera vibrios identified in Ukraine in 1994 and 2011 such as a typical toxigenic biovar El Tor (V cholerae Ο1, El Tor, Ogawa, Hly-, ctxA+, tcpA+) contain genes ofthe classic cholera vibrio in their genome and are genetically altered (hybrid) variants of cholera vibrio biovar El Tor producing enterotoxin CTI and having increased virulence, that was clinically manifested in predominance of severe forms of cholera in Mariupol of Donetsk region in 2011. Genome sequences ofthe 4 studied strains were deposited into the international database DDBJ/EMBL/GenBank. CONCLUSION: The studied isolates were established to belong to a clade of strains associated with cholera outbreaks in Haiti and Asian continent, from where genetically altered strains of cholera vibrios biovar El Tor were introduced to Haiti in 2010, based on results of comparison of genomic sequences of the studied strains with genomes of V cholera strains from the international database GenBank.

[EPIDEMIOLOGIC SITUATION BY NATURAL-FOCI INFECTIONS IN THE CRIMEA FEDERAL DISTRICT IN 2014-2015].

AIM: Analysis of epidemic manifestations of natural-foci infections (NFI), clarification of spectrum of their causative agents, determination of epizootic activity of natural foci in the Crimea Federal District (KFD). MATERIALS AND METHODS: Epizootologic examination of 10 administrative districts of KDF was carried out. 291 pools (2705 specimens) of ixodes ticks and 283 samples of organs of small mammals were studied by PCRmethod for the presence of DNA/RNA of causative agents of a number of NFI. RESULTS: Morbidity by NFI in KFD was registered by 6 nosologies: Lyme borreliosis, Marseilles fever, leptospirosis; tularemia, intestine yersiniosis and tick-borne viral encephalitis, wherein, transmissive infections made up 91.6%. Circulation of causative agents of Crimea hemorrhagic fever, Q fever, group of tick-borne spotted fever, Lyme borreliosis, human granulocytic anaplasmosis, human monocytic ehrlichiosis, hemorrhagic fever with renal syndrome, West Nile fever, tularemia and leptospirosis was established. CONCLUSION: Due to activity of natural foci of NFI further monitoring of epidemiologic and epizootologic manifestations of these infections in the Crimea, including using genetic methods of analysis, is necessary for ensuring sanitary-epidemiologic welfare of KFD population.

A NEW GENETIC VARIANT OF THE CRIMEAN-CONGO HEMORRHAGIC FEVER VIRUS ISOLATED IN CRIMEA.

This work represents the results of the genetic identification of the Crimean-Congo hemorrhagic fever virus (CCHF virus) strains isolated in the Crimean Federal District in conducting the epidemiological survey of the imported case of CCHF from Crimea in 2015. One sample of the serum from a patient and 61 pools (506 specimens) of ticks collected during the epizootiological survey of 6 administrative districts of the Crimean Federal District were tested using PCR for the presence of the CCHF virus RNA. RNA of the CCHF virus was detected in serum from a patient and 10 samples of ticks. The genetic identification of the CCHF virus was performed by sequencing the virus genome S-, M-, and L-segments. The result of the molecular-genetic analysis revealed a high degree of identity between the samples of the CCHF virus in human serum and three samples of ticks and their belonging to a new genetic Crimea subclade (Vd) of the genotype Europe 1. Whole genome sequencing of two samples of CCHF virus belonging to the Crimea subgroup (Vd) was performed. CCHF virus variants of the Crimea subclade (Vd) of the Europe- lgenotype were described for the first time. These variants were endemic to the territory of the Crimean peninsula.

[PROCUREMENT OF SANITARY AND EPIDEMIOLOGICAL WELFARE IN THE REGION OF-CITY-RESORT SOCHI FOR DANGEROUS AND NATURAL-FOCI INFECTIOUS DIS- EASES IN 2015].

AIM: Analysis ofresults of epidemiologic monitoring especially dangerous, natural-foci and other infectious diseases, asill as epizootologic activity of natural foci of infection on,the terri- tory, of city-resort Sochi. MATERIALS AND METHODS: Laboratory studies of 820. samples by PCR; im- mune- and bacteriologic methods were carried out, among those 344 - clinical material, 12 - water from open bodies and 321 - field material. Molecular-genetic identification of-143 strains of Vibrio-cholerae, isolated from open water bodies of city-resort Sochi, was-carried out,. RESULTS: Circulation of causative agents of Q fever, tularemia and hemorrhagic fever with renal syndrome of Dobrava-Adler genotypes was established, -as well as rickettsia of tick spotted fever group Predomination of rotaviruses (70.9%) was detected during study of samples of clinical material in etiologic structure of sporadically emerging acute intestine infections. Relatively high temperature values of river water during summer months (from 18, to 30 C), exit of sulfide mineral waters into the riverbed, that create-alkaline medium, seasonal reduction of river debit and speed of water current facilitated sustenance of contamination of water of Agura river by V.cholerae. CONCLUSION: Activity of natural foci of the infectious disease and contamination of Agura iver by V.cholerae Ο1, (atoxigenic) gives evidence on the necessity of constant control of epidemiologic situation by dangerous and natural-foci infectious diseases on the territory of city-resort Sochi, as well as study regional features, of their causative agents, including using genetic methods.

[Modern methods application of genotyping of infectious diseases pathogens in the context of operational work of specialized anti-epidemic team during the XXII Olympic Winter Games and XI Paralympic Winter Games].

This paper considers the experience of genotyping and sequencing technologies in laboratories of specialized anti-epidemic team (SAET) during the XXII Olympic Winter Games and XI Paralympic Winter Games of 2014 in Sochi. The work carried out during the pre-Olympic period on performance of readiness by SAET for these studies is analyzed. The results of genotyping strains of pathogens during the Olympic Games are presented. A conclusion about the effectiveness of the use of molecular genetic techniques in terms of SAET is made.