The Development And Application Of Rapid Detection Methods For Hog Cholera And Porcine Reproductive And Respiratory Syndrome

Hog Cholera(HC) is a highly infectious viral disease caused by the agent of Hog Cholera virus which is the member of genus pestivirus, the family flaviviridae. Porcine reproductive and respiratory syndrome(PRRS), aroused by the PRRS virus, is a kind of highly contagious disease that is a great danger to swine cultivation industry. In recent years,those two diseases were take great side effects to swine industry. Due to establishing rapid,specific and convienent diagnosis technology is critical to control and prevent two diseases, this research using molecular biology and immunology developed few methods for detection of HC and PRRS and it have greatly significance to swine cultivation industry.According to HC and PRRS virus conservative sequence,this study was designed two pairs of specific HC and PRRS primers,and developed Multiplex RT-PCR which could detect HC and PRRS simultaneously. one pair of specific primer was designed according to the HC virus 5'NTR nucleotide sequence downloaded from Genebank,moreover,established SYBR GreenI fluorescent real-time RT-PCR method for detection of HC virus. one pair of specific primer was designed according to the PRRS M gene nucleotide seqence downloaded from Genebank,moreover,established SYBR GreenI fluorescent real-time RT-PCR method for detection of PRRS virus.According to the immunology property of HC and PRRS virus, based on HC and PRRS single dot immunogold filtration assay (DIGFA),this study developed the double DIGFA that could detect HC and PRRS simultaneously.Establishing those methods not only provide the theoretical foundation and method for clinical detection of two diseases,but also is significant to conduct further study for the prevention and clinical detection of two diseases.