Morozov Vladimir A, Plotzki Elena, Rotem Avi, Barkai Uriel, Denner Joachim
HIV and other Retroviruses, Robert Koch Institute, Berlin, Germany.
Beta-O2 Technologies Ltd., Rosh-Haain, Israel.
Xenotransplantation. 2016 Nov;23(6):490-496. doi: 10.1111/xen.12265. Epub 2016 Sep 11.
To prevent transmission of zoonotic microorganisms from pig transplants to human recipients when performing xenotransplantation using pig cells, tissues, or organs, donor pigs have to be carefully characterized. Göttingen minipigs (GöMP) are often used for various biomedical investigations and are well characterized concerning the presence of numerous bacteria, fungi, viruses, and parasites. Recently, we studied the prevalence and expression of porcine endogenous retroviruses and the prevalence of hepatitis E virus (HEV) in GöMP. Here, we studied the presence of the porcine cytomegalovirus (PCMV) and porcine lymphotropic herpesviruses (PLHV) and extended testing for hepatitis E virus (HEV).
PCR, nested PCR, real-time PCR, real-time RT-PCR, and Western blot analyses were used to estimate the prevalence of PCMV, PLHV-1, PLHV-2, PLHV-3, and HEV.
Using different PCR methods, and different source materials, PCMV was found in 10 of 26 adult GöMP, which had been derived originally by cesarean section and kept under specified pathogen-free conditions. Only highly sensitive methods gave positive results, not methods of lower sensitivity. The virus load in all positive animals was low (<100-200 copies per mL). PLHV-1, PLHV-2, and PLHV-3 were not detected by PCR; however, an anti-PLHV immune response was found in one of 10 animals tested by Western blot analyses. HEV was detected by RT-PCR in two of nine tested animals, but no anti-HEV immune response was observed.
Using highly sensitive methods, PCMV, HEV, and PLHV were found in some GöMP, suggesting that these viruses may be introduced through the placenta. The results show that highly sensitive methods are required to characterize pigs to be used for xenotransplantation to prevent virus transmission.
在使用猪细胞、组织或器官进行异种移植时，为防止人畜共患病微生物从猪移植物传播给人类受体，必须对供体猪进行仔细的特征鉴定。哥廷根小型猪（GöMP）常用于各种生物医学研究，并且在众多细菌、真菌、病毒和寄生虫的存在方面具有详细的特征描述。最近，我们研究了猪内源性逆转录病毒的流行情况和表达以及哥廷根小型猪中戊型肝炎病毒（HEV）的流行情况。在此，我们研究了猪巨细胞病毒（PCMV）和猪嗜淋巴细胞疱疹病毒（PLHV）的存在情况，并扩展了对戊型肝炎病毒（HEV）的检测。
采用聚合酶链反应（PCR）、巢式PCR、实时PCR、实时逆转录PCR和蛋白质印迹分析来评估PCMV、PLHV-1、PLHV-2、PLHV-3和HEV的流行情况。
使用不同的PCR方法和不同的来源材料，在26只成年哥廷根小型猪中的10只中发现了PCMV，这些猪最初通过剖宫产获得并饲养在特定病原体-free条件下。只有高灵敏度的方法得出阳性结果，低灵敏度的方法未得出阳性结果。所有阳性动物中的病毒载量都很低（每毫升<100-200拷贝）。通过PCR未检测到PLHV-1、PLHV-2和PLHV-3；然而，在通过蛋白质印迹分析检测的10只动物中的1只中发现了抗PLHV免疫反应。通过逆转录PCR在9只检测动物中的2只中检测到HEV，但未观察到抗HEV免疫反应。
使用高灵敏度方法，在一些哥廷根小型猪中发现了PCMV、HEV和PLHV，表明这些病毒可能通过胎盘传播。结果表明，需要使用高灵敏度方法对用于异种移植的猪进行特征鉴定，以防止病毒传播。
