THE OPTIMIZATION PROCESS OF MULTIPLEX PCR TO DETECT AND IDENTIFY THE GENOTYPES OF AFRICAN SWINE FEVER VIRUS IN VIETNAM

Abstract

Multiplex PCR is a widely used molecular biology method in many studies because of its superiority. A reaction that amplifies multiple DNA sequences in a single PCR reaction. Therefore, we have studied and applied the molecular biotechnology technique of Multiplex PCR to detect the presence of the ASF virus and identify which genotype is circulating in Vietnam. The objective is to design and optimize the multiplex PCR reaction as the basic kit for detecting and genotyping the ASF virus in Vietnam. The results showed that the multiplex PCR reaction was successfully designed and optimized, including the concentration of primers Buffer 1X; dNTP 0.5 µM; 1.25 mM MgCl2; 0.05 U/µl Taq DNA polymerase, primers 0.1 – 0.4 µM (p72-F, p72-R, p72-SNP, p72-g01, p72-g02, p72-g08, p72-g09, p72-g10, p72-g23); ≥ 150 pmol/µl DNA standard. Heat cycle is 95^oC - 5 minutes; (95^oC – 30 sec; 62^oC – 15 sec; 72^oC – 30 sec) x 30 cycles; 72^oC - 7 minutes. This result can be used to develop a kit to detect and identify the ASF virus, which is meant for use in the Vietnamese livestock industry.