METHOD FOR INDIRECT DETERMINATION OF THE COMPLETENESS OF FMD VIRUS ANTIGEN INACTIVATION IN VACCINE RAW MATERIALS USING REVERSE TRANSCRIPTASE POLYMERASE CHAIN REACTION IN REAL TIME WITH AMPLIFICATION OF LARGE-SIZED FRAGMENT

摘要

FIELD: biotechnology.;SUBSTANCE: invention relates to the field of biotechnology, molecular diagnostics and production of FMD vaccines. Described is a method for indirectly determining the completeness of FMD virus antigen inactivation in raw materials for a vaccine using reverse transcriptase polymerase chain reaction in real time (RT-PCR-RT) with amplification of a large-sized fragment. The study of the completeness of inactivation of the FMD virus antigen in the raw material for inactivated FMD vaccines is determined by elution of viral RNA, assessment of the purity and concentration of RNA using spectral scanning and the formula С_RNA = 40.5 × OD₂₆₀ - 0.7037, reverse transcriptase polymerase chain reaction in real time, with the help of which an intact 7279 nt region of the FMD virus genome is detected. in the range from 5'-NTR to 3D-gene inclusive, the presence of an exponent and, thus, determine the presence of a virulent FMD virus, or reveal the absence of an amplicon of the required size, the absence of an exponent and, thereby, prove the absence of a virulent FMD virus in raw materials for the production of FMD inactivated vaccines. ;EFFECT: method is economical, it allows simultaneously examining several tens of samples, and the analysis time can be reduced to 9-10 hours, is characterized by high diagnostic sensitivity - 99.44%, specificity - 100% and overall accuracy - 99.72%.;8 cl, 6 dwg, 9 tbl, 6 ex.