The Establishment And Multi-application Of PPA-ELISA And Nest-PCR Of SARS Associated CoV.

The pathogen of Severe Acute Respiratory Syndrome(SARS) is Corona Virus, For investigating the truth about Corona virus in animals, and find the homologous with SARS. CoV, a essay have been establish that based on the characters that SPA can combine the IgG in the serum of many kinds of animals, indirect PPA-ELISA. To detect antibody of SARS virus in the serum of many animals was established. Two pairs of primer was designed according to the partly polymerase gene of SARS associated corona virus sequences published in Genbank. 146 samples from 11 kinds of animals were detected by nest-PCR., Then PCR product was sequenced. Phylogenetic analysis was done on the amplified region .And the relationship between Corona virus for the animals and SARS. CoV was analyzed.The results indicated that SPA can combine with many mammals IgG in the serum well, and determined the best time of the antigen combined antibody reaction HRP-SPA commeded antibody, At the same time, we made the method of PPA-LEISA which detected the SARS antibody dedicational.The result of amplifying of polymerase gene of SARS associated corona virus and sequence analyze showed that there were high homology between the samples from human and animals, around 100%~95. 8% in nucleotide acid.By detect the sample of the same animals using PPA-ELISA and nest-PCR separated, the accordance rate of the two methods is around 60%~70%, it indicated that it' s valuable for the detection of Corona virus in animals by using the two assay.