Open AccessMETHOD FOR FMD VIRUS 146S COMPONENT CONCENTRATION DETERMINATION WITH REAL-TIME REVERSE TRANSCRIPTION – POLYMERASE CHAIN REACTION IN VACCINE RAW MATERIALS
Author(s) -
М. И. Доронин,
А. М. Тимина,
D. A. Lozovoy,
В. А. Стариков,
Д. В. Михалишин,
Н. Н. Медведева,
А. В. Борисов
Publication year - 2018
Publication title -
veterinariâ segodnâ
Language(s) - English
Resource type - Journals
eISSN - 2658-6959
pISSN - 2304-196X
DOI - 10.29326/2304-196x-2018-2-25-31-35
Subject(s) - virology , reverse transcription polymerase chain reaction , real time polymerase chain reaction , reverse transcriptase , polymerase chain reaction , complement fixation test , antigen , virus , biology , immunology , serology , antibody , gene , biochemistry , messenger rna
Method for determination of foot-and-mouth disease (FMD) virus 146S component concentration with real-time reverse transcription – polymerase chain reaction (rtRT-PCR) in vaccine raw materials is developed. Negative regression model of С 146S = (30.269 – C t )/4.155 type allowing determination of FMDV 146S particle concentrations based on the amplification threshold cycle values (C t ) is proposed. It has been experimentally proven that the quantity of the 146S component determined by the real-time RT-PCR method using developed regression model and contained in the inoculation dose of FMD vaccine confers protection to the vaccinated animals against generalized FMD of A, O, Asia-1 types. rtRT-PCR method is highly sensitive and allows rapid and highly reliable estimation of the 146S antigen concentration in FMD vaccine. The method for 146S particle quantity determination by real-time RT-PCR using the regression model is reliable and demonstrates high correlation (95.5–99.0%) with the complement fixation test results.