DEVELOPMENT AND STUDY OF THE PERFORMANCE OF THE REAGENT KIT FOR THE IDENTIFICATION OF VIBRIO CHOLERAE AND VIBRIO PARAHAEMOLYTICUS BY MULTILOCUS ALLELE-SPECIFIC PCR IN CLINICAL TRIALS

Abstract

The aim of the study was to develop, conduct clinical trials and evaluate the performance of the "Reagent kit for identification of V. cholerae and V. parahaemolyticus by multilocus allele-specific PCR" ("Vibrio-screen FL") for the state registration of the specified medical device. Materials and methods. The results of clinical trials of the reagent kit "Vibrio-screen FL" that has been developed to optimize the means and methods of identification and differentiation of pathogens of particularly dangerous infections are presented in the article. 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). To conduct clinical trials clinical strains of microorganisms Vibrio spp. and other microorganism species were used as target analytes. Research methods included bacteriological method and multilocus PCR . 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, there were no non-specific reactions with DNA samples of other microorganism species. A medical device for in vitro diagnostics the reagent kit 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 the method of multilocus allele-specific polymerase chain reaction with hybridization-fluorescent detection. The registration certificate № РЗН 2021/13360 from 08.02.2021 was received. By order of Roszdravnadzordated 08.02.2021 No. 1041, a reagent kit Vibrio-screen FL by the Rostov-on-Don Plague Control Researsh Institute 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 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, which allows it to be used in clinical laboratory diagnostics, for epidemiological analysis.