CAJ | 학술논문

目的：建立同时检测口蹄疫病毒、水泡性口炎病毒和猪水泡病病毒的多重荧光RT-P C R检测方法。方法根据口蹄疫病毒3D蛋白编码基因、水泡性口炎病毒N蛋白编码基因和猪水泡病病毒VP1蛋白编码基因的高保守区设计特异性引物和探针，对3种动物病毒进行多重荧光定量RT-PCR扩增。结果经过扩增，可以同时检测口蹄疫病毒、水泡性口炎病毒和猪水泡病病毒，而其他参试病原均无扩增信号，显示其良好的特异性。对口蹄疫病毒、水泡性口炎病毒、猪水泡病病毒的最低检测限分别达到101、102、102个质粒拷贝浓度。结论本方法灵敏度高，特异性良好，可实现多种病毒混合感染的同时检测。
목적：건립동시검측구제역병독、수포성구염병독화저수포병병독적다중형광RT-P C R검측방법。방법근거구제역병독3D단백편마기인、수포성구염병독N단백편마기인화저수포병병독VP1단백편마기인적고보수구설계특이성인물화탐침，대3충동물병독진행다중형광정량RT-PCR확증。결과경과확증，가이동시검측구제역병독、수포성구염병독화저수포병병독，이기타삼시병원균무확증신호，현시기량호적특이성。대구제역병독、수포성구염병독、저수포병병독적최저검측한분별체도101、102、102개질립고패농도。결론본방법령민도고，특이성량호，가실현다충병독혼합감염적동시검측。
ABSTRACT:Objective To establish a multiplex real-time RT-PCR method for simultaneous detection of foot and mouth disease virus, vesicular stomatitis virus and swine vesicular disease virus.Methods According to the gene encoding 3D protein of foot-and-mouth disease virus, vesicular stomatitis virus N protein coding gene and swine vesicular disease virus VP1 protein encoding gene, specific primers and probe were designed, and 3 kinds of animal viruses were amplified by multiplex real-time RT-PCR.Results The FMDV, VSV and SVDV could be simultaneously detected after amplification, and other tested pathogens were no amplification signal, displaying the good specificity. The minimum detection limits of foot and mouth disease virus, vesicular stomatitis virus and swine vesicular disease virus were 101, 102, and 102 plasmid copy concentration, respectively.Conclusion This method has a good specificity and sensitivity for simultaneous detection of FMDV, VSV and SVDV at the same time.