Multiplex PCR For Simultaneous Detection And Differentiation Of Goatpox, Sheeppox, Orf And Pester Des Petits Ruminants Viruses

Goatpox virus, sheep pox virus, orf virus and peste des petits ruminants virus can infect goats and sheep single or multiple infections. For the rapid detection and identification of any kinds of viral diseases, the establishment of a multiplex PCR. According to the GenBank database has included goat pox virus, sheeppox virus,orf virus and peste des petits ruminants virus gene sequences, strict adherence to the principles of primer design, use Primer Primer 5.0 designed four pairs of primers for multiplex PCR. In this study, the optimal screening by conventional PCR amplification reaction conditions and the optimal annealing temperature, the results showed that the optimum conditions were:denaturation temperature of 94℃, annealing temperature of 53℃,35 times the number of cycles. Test goat pox virus% sheep pox virus、orf virus and peste des petits ruminants virus PCR product length were 152bp,246bp,355bp,538bp, with the expected length of the amplified product design primers are consistent; And this study of PCR result was negative for bovine viral diarrhea-mucosal disease virus, swine fever virus and infectious bovine rhinotracheitis virus, indicated that the test has a high specificity; The PCR reaction system could detect the goat pox virus, sheep pox virus, orf virus and peste des petits ruminants virus minimum amount were 0.05ng、8.36ng、0.8ng、 1.3ng.The results showed that, the characteristics of this experiment is rapid, high authority and specificity. It can be used for differential diagnosis in clinical analysis about goat pox, sheep pox, orf and peste des petits ruminants.