FIELD: biotechnology.
SUBSTANCE: method of indirect determination of the completeness of inactivation of the rabies virus antigen using reverse transcriptase polymerase chain reaction (hereinafter - RT-PCR) followed by electrophoresis of amplicons in agarose gel is described. The completeness of inactivation of the rabies virus antigen is determined by RT-PCR using 5 systems of original primers (Shift Oligo-NR1069 (for the site of the highly conserved gene N of the rabies virus); PF1807-MR3020 (for the site that captures the genes P and M of the rabies virus); GF3471-ΨR5520 (for the site of the variable fragment of the G gene and the Ψ-area of the rabies virus); LF5499-LR6290 and LF8085-LR9954 (for the sites of the L gene of the rabies virus carrying information about RNA-dependent DNA polymerase)). Detection of the results of completeness of inactivation of the rabies virus is carried out in an ultraviolet light flux at a wavelength of 312 nm after horizontal electrophoresis in 1.5% agarose gel with the addition of 0.0005% ethidium bromide. In the presence of fluorescent amplicons obtained in PCR using 5 systems of original primers, the sample is virulent; in the absence of luminous PCR products in all 5 systems of primers, the sample is inactivated.
EFFECT: present invention makes it possible to assess quickly and with a high degree of confidence the completeness of inactivation of the rabies virus antigen using a reverse transcriptase polymerase chain reaction followed by electrophoresis of amplicons in an agarose gel.
8 cl, 8 tbl, 4 ex, 7 dwg
