Diagnosis Of BVDV And Dividing Type Studying Using RT-PCR

Bovine viral diarrhea/mucosal disease (BVD/MD), abbreviation called BVD or MD, which caused by Bovine viral diarrhea-mucosal disease virus, represented the prototype member of the pestivirus genus, family Flaviviridae. Bovine infected by BVDV showed diarrhea, acute and chronic mucosal diaease, persistent infection and immunotolerance, immunosupression, pregnant cow abortion, dead fetus and abnormal fetus. BVDV was a worldwide pathogen in cattle which had not been controlled by classical vaccination,and had seriously endangered the cattle herds. Although BVDV have only one serum type, it can be divided into two genotypes according to 5'untranslated region(5'-UTR), BVDVâ… and BVDVâ…¡. Meanwhile, according to cytopathic effect BVDV can be divided into two biotypes, CP and NCP. This Research set up a measure which can distinguish genetype and biotype using RT-PCR of BVDV, and it is foundation of fast Diagnose for BVD. According to already published BVDV genome sequence in GenBank, give consideration to the clear array difference of different serum type of BVDV and difference among HCV and BDV array. 5'-UTR of BVDV is very high conservative among each genesome, which is the main differenence of BVDVâ… and BVDVâ…¡. So it choose at 5'-UTRof BVDV design distinguished genotype primers of BVDV. Whether the biotype conversion of BVDV involves and has a other source RNA that is inserted mainly, So determine the biotype quotes things to choose to be inserted in some conservative districts of both ends in the location in RNA. Utilize RT-PCR method, clone and receive the purpose passage, check sequence, analyse the comparative homology, the result, with expecting to conform. Sensitiveness test, utilize Reed-Muench method examine BVDV cell train malicious TCID50, after ten times of gradient dilute, draw RNA separately , increase PCR, the result of the test indicates that can be measured to one TCID50. Peculiar test, choose foot-and-mouth disease and virus , ox's nose bronchitis virus , bubble stomatitis virus with similar clinical symptom, have serum learn swine fever virus that react alternately , draw RNA (DNA ) separately , amplification by PCR, three pair primers can only regard cDNA that RNA of BVDV duplicates template instead as , is it in conformity with each experimental design the purpose passage to happen to increase, and the increase result of contrasting the poisonous Corporation is negative . Prove RT-PCR detection method is peculiar very high. The animal comes back to test, it can utilize RT-PCR in the whole blood their and infect virus early nose is it reach BVDV to be positive to measure among the liquid on bovine to inoculate. The above-mentioned results show RT-PCR detection method set up in this research, not only can diagnose BVD accurately fast but also utilize different guiding genotype and living beings type that the thing can distinguish BVDV. Thus offer the strong diagnostic tool to the measuring of this disease for the cattle farm.