Porcine Endogenous Retrovirus In Porcine-derived Medical Devices Detection Method And Albumin Biological Glue Research,

This paper includes two parts. In the first part, the objective is to design a real-time PCR test to detect porcine endogenous retrovirus (PERV) RNA in the medical devices in which the materials derived from porcine. The primers and probe for PCR and real-time PCR were designed by the oligo software and Primer Express software based on the highly conservative regions, which were found by the DNASTAR software. The target region was amplified by PCR from the genomic DNA of the PK-15 cells and cloned into pGEM-T vector. The presence and orientation of the inserts were confirmed by restriction enzyme digestion and DNA sequencing. The cloned vectors were transcribed into RNA and the concentration of the RNA was determined by spectrophotometer. The standard curve of the RNA was established and the samples were detected. Real-time PCR assay had a sensitivity of 1000 RNA copies per reaction, with a linear range of detection between 10~3 and 10~7 RNA copies. The intra- and inter-runs coefficient variations of threshold cycle (Ct) values were less than 1.44% and 2.97%, respectively. The results of the samples were all positive and the inter-assay variation was less than 1.93%. The result of reverse transcriptase (RT) assay was also positive. The conclusion was that Real-time PCR not only had fast throughput capability but also had highly sensitivity, highly specificity, highly reproducibility and was suitable for the detection of medical devices in which the materials derived from porcine. This assay had a broad application for viral safety evaluation of these products. The samples detected here were all used in vivo, more attention should be paid on the viral safety of these products.In the second part, the objective was to prepare a kind of surgical adhesive in which one of the ingredients was albumin. The albumin was purified from pocine plasma by hot ethanol denaturalization method in which albumin was protected by sodium caprylate while other proteins were denaturalized and deposited. This...