METHOD FOR INDICATION AND DIFFERENTIATION OF VACCINE STRAINS AND FIELD ISOLATES OF CLASSIC HOG CHOLERA VIRUS

摘要

FIELD: veterinary virology, molecular diagnostics. SUBSTANCE: method includes the synthesis of external universal and internal CHCV-specific oligonucleotide primers, isolation of viral RNA out of vaccine preparations or pathological material, synthesis of cDNA on the matrix of viral RNA in reverse transcription reaction. Then comes the first stage of amplification of viral genome fragment in polymerase chain reaction by using external universal oligonucleotide primers. The second stage of amplification should be carried out in polymerase chain reaction by using internal CHCV-specific oligonucleotide primers. Then one should conduct the restriction of obtained fragments of cDNA with endonuclease Taq I enzyme. Differentiation of vaccine strains and field isolates of CHCV should be carried out based upon the data on the quantity and weight of obtained restriction fragments after electrophoresis in a polyacrylamide gel. The method enables to obtain increased specificity and sensitivity of the technique conducted. EFFECT: higher efficiency. 1 dwg, 2 ex