Establishment Of Three Different Methods For Detecting PRRSV

Porcine reproductive and respiratory syndrome virus causes pigs to be reproductive failure,abortions,stillborn fetuses,mummified fetuses and weak-born pigs and significant economic losses. In recent years, a new highly pathogenic viruses appears in China. To detect this new virus, a series of methods were necessary to be established. The methods of RT-PCR,RT-LAMP and Real-Time was used to detect the mutational virus.Highly pathogenic PRRSV was research object. Many gene sequences of highly pathogenic PRRSV and normal PRRSV were downloaded from NCBI. The structural features was analyzed between them. Nsp2 gene and ORF7 gene was to be the detective genes. Primers and probes were designed to detect PRRSV by the methods of RT-PCR,RT-LAMPå’ŒReal-Time RT-PCR.PRRSV was inoculated in Marc-145 cells and then observed CPE. Diseased cells frozened and thawed for 3 times, and then subcultured virus. Culture medium was used for the tests of RT-PCR,RT-LAMP and Real-Time. The results revealed highly pathogenic PRRSV lacked 87bp contrasted with normal PRRSV,but ORF7 gene was conserved. These methods were established for deferent detective conditions,and facilitated to detect PRRSV.