METHOD AND SET FOR DETECTION OF AFRICAN PLAGUE PIG VIRUS DNA BY METHOD OF POLYMERASE CHAIN REACTION Russian patent published in 1999 - IPC

FIELD: molecular biology, veterinary virology, genetic engineering. SUBSTANCE: invention relates to a method and set for detection of pathogen DNA of African plague pig in culture of infected cells and samples of pathological material from pigs. Method involves amplification of DNA-fragment (size is 1144 nucleotide pairs) with primers N1 and N3 and fragment containing 562 nucleotide pairs (repeated amplification) with primers N2 and N3 containing an unique site for restriction endonuclease Pst I. Method involves the use of primers showing complementarity to gene of basic structural protein Vp 72 located in conservative region of African plague pig virus. Set has plastic tubes containing: specific primers (N1, N2, N3), thermostable enzyme Taq-polymerase, 10-fold buffer for reaction, a mixture of deoxynucleotide tri-phosphates, positive control - recombinant plasmid containing gene that encodes protein Vp 72. Method ensures to detect up to 10 fentograms DNA of African plague pig virus or 10 particles in 1 ml infectious virus in different materials including pig organ samples with chronic and latent forms of disease course. Time analysis is 4 h. EFFECT: improved method of analysis, enhanced effectiveness of method, accelerated procedure. 3 cl, 3 tbl, 1 ex