The Establishment Of The Method To Detect The Porcine Parvovirus Potentially Contaminated In The Biological Products For Human Use From The Swine

To ensure the security and the quality of the porcine parvovirus in the biological products for human use from the swine, it is necessary to detect the related virus potentially contaminated in the products. The purpose of this research work is to establish the method involved in the molecular biology and the serology to detect the PPV in the related biological products.By searching the gene sequence of the PPV, other parvovirus and virus growthing in the swine which published in the GeneBank , analyzing the difference among these sequence and using the Primer primier5.0 and Oligo6.0 according to the primer designing principle we designed a pair of idio-primer to aim directly at the PPV, which can be used to amplify a fragment in 531 bp length. The optimal conditions of the amplification was determined by the square matrix titrating method. PCR product was confirmed by the dot-blot-hybrization with the oligonucleotide probe, conjucting PCR product with the pGEM-T-easy vector and then sequencing the product. Diluted the template in 10 times extracted from the virus culture whose TCID₅₀ was titrated as 10^-3.6, then amplifying the diluted template respectively, we can detect the minimum TCID₅₀ of 0.398. With the PCR detection, we have detected the swine organ from slaughter houses in BJ and the PPV and swine kidney conjionedly contaminating PK-15 model.Five BALB/c mice(female, 6-8 weeks old) were immunized with the purified...