Yang Yang, Qin Xiaodong, Zhang Wei, Li Yanmin, Zhang Zhidong
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China.
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Xujiaping 1, Lanzhou, 730046, Gansu, China.
Mol Cell Probes. 2016 Oct;30(5):300-305. doi: 10.1016/j.mcp.2016.08.011. Epub 2016 Sep 1.
Porcine parvovirus (PPV) is a major cause of swine reproductive failure and reported in many countries worldwide. Recombinase polymerase amplification (RPA) assays using a real-time fluorescent detection (PPV real-time RPA assay) and a lateral flow dipstick (PPV RPA LFD assay) were developed targeting PPV NS1 gene. The detection limit of PPV real-time RPA assay was 300 copies per reaction within 9 min at 38 °C, while the RPA LFD assay has a detection limit of 400 copies per reaction in less than 20 min at 38 °C. In both assays, there were no cross-reactions with porcine circovirus type 2, pseudorabies virus, porcine reproductive and respiratory syndrome virus, classical swine fever virus, and foot-and-mouth disease virus. Based on a total of 128 clinical samples examined, the sensitivity and the specificity of the developed RPA assays for identification of PPV was 94.4% and 100%, respectively, when compared to real-time (qPCR) assay. Therefore, the RPA assay provides a rapid, sensitive and specific alternative for PPV detection.
猪细小病毒（PPV）是导致猪繁殖障碍的主要原因，在世界许多国家均有报道。针对PPV NS1基因开发了使用实时荧光检测的重组酶聚合酶扩增（RPA）检测方法（PPV实时RPA检测）和侧向流动试纸条检测方法（PPV RPA LFD检测）。PPV实时RPA检测在38℃下9分钟内每个反应的检测限为300拷贝，而RPA LFD检测在38℃下不到20分钟内每个反应的检测限为400拷贝。在这两种检测方法中，与猪圆环病毒2型、伪狂犬病病毒、猪繁殖与呼吸综合征病毒、经典猪瘟病毒和口蹄疫病毒均无交叉反应。基于总共检测的128份临床样本，与实时（qPCR）检测相比，所开发的RPA检测方法鉴定PPV的灵敏度和特异性分别为94.4%和100%。因此，RPA检测为PPV检测提供了一种快速、灵敏且特异的替代方法。
