猪内源性逆转录病毒检测的三重PCR方法的建立及其在五指山猪组织样品检测中的初步应用

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中图分类号：S854.43 文献标志码：A 文章编号：0366-6964（2025）07-3548-07

Abstract：This study aims to establish a preliminary triplex PCR assay capable of simultaneously detecting porcine endogenous retroviruses （PERV） in the pig genome and differentiating the three PERV subtypes： PERV-A， PERV-B，and PERV-C. Specific primers were designed based on the envelope （env） gene of different PERV subtypes. The method was validated for specificity， sensitivity and reproducibility after optimization of the reaction conditions and the reaction system. The results showed that the triplex PCR method specifically amplified all three PERV subtypes. The detection limits for the plasmid standards of PERV-A，PERV-B，and PERV-C were 1.50×102 ， 1.99×103 ，and 1.57×102 copies·μL-1 ，respectively. A total of 103 tissue samples were tested using the triplex PCR method and compared with the conventional single PCR method. The positive rates were 100% （103/103）for PERV-A， 100% （103/103）for PERV-B and 54% （56/lo3） for PERV-C. The compliance rates for the two methods were PERV-A，PERV-B，and PERV-C were 100% ， 100% ，and 96.6% ，respectively. This study successfully developed a highly specific， sensitive， rapid，and convenient triplex PCR assay， which can be used for the simultaneous detection and differential diagnosis of the three PERV subtypes，and provides technical support for the screening of PERV-free porcine-derived donor organs.

Keywords： porcine endogenous retrovirus;env gene; triple PCR method; tissue samples ∗ Corresponding author： LIU Guangliang，E-mail： LiuGuangliangO1@caas. cn

随着医学免疫学和移植免疫学的不断进步，器官移植在治疗患者终末期疾病中发挥着重要的作用，但人类移植器官供体存在严重短缺现象，因此，异种器官移植得到广泛关注，有望成为未来救助需要器官移植病人的有效手段之一。（剩余9428字）