Yang Haowen, Liu Ming, Zhou Bingcong, Deng Yan, He Nongyue, Jiang Hesheng, Guo Yafen, Lan Ganqiu, Jiang Qinyang, Yang Xiurong, Li Zhiyang
J Nanosci Nanotechnol. 2016 Jun;16(6):6505-10. doi: 10.1166/jnn.2016.10841.
Chinese Bama minipigs could be potential donors for the supply of xenografts because they are genetically stable, highly inbred, and inexpensive. However, porcine endogenous retrovirus (PERV) is commonly integrated in pig genomes and could cause a cross-species infection by xenotransplantation. For screening out the pigs with low copy numbers of PERV proviruses, we have developed a novel semiquantitative analysis approach based on magnetic nanoparticles (MNPs) and chemiluminescence (CL) for estimating relative copy numbers (RCNs) of PERV proviruses in Chinese Bama minipigs. The CL intensities of PERV proviruses and the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were respectively determined with this method, and the RCNs of PERV proviruses were calculated by the equation: RCN of PERV provirus = CL intensity of PERV provirus/CL intensity of GAPDH. The results showed that PERVs were integrated in the genomes of Bama minipigs at different copy numbers, and the copy numbers of PERV-C subtype were greatly low. Two Bama minipigs with low copy numbers of PERV proviruses were detected out and could be considered as xenograft donor candidates. Although only semiquantitation can be achieved, this approach has potential for screening out safe and suitable pig donors for xenotransplantation.
中国巴马小型猪可能是异种移植供体的潜在来源，因为它们基因稳定、高度近亲繁殖且成本低廉。然而，猪内源性逆转录病毒（PERV）通常整合在猪基因组中，可能通过异种移植导致跨物种感染。为了筛选出PERV前病毒拷贝数低的猪，我们开发了一种基于磁性纳米颗粒（MNPs）和化学发光（CL）的新型半定量分析方法，用于估计中国巴马小型猪中PERV前病毒的相对拷贝数（RCNs）。用该方法分别测定PERV前病毒和看家基因甘油醛-3-磷酸脱氢酶（GAPDH）的CL强度，并通过以下公式计算PERV前病毒的RCNs：PERV前病毒的RCN = PERV前病毒的CL强度/GAPDH的CL强度。结果表明，PERV以不同拷贝数整合在巴马小型猪的基因组中，且PERV-C亚型的拷贝数极低。检测出两头PERV前病毒拷贝数低的巴马小型猪，可被视为异种移植供体候选者。尽管只能实现半定量，但该方法有潜力筛选出安全合适的异种移植猪供体。
