水貂阿留申病毒环介导等温扩增（LAMP）检测方法的建立

摘要

为建立一种简便、快速、特异的水貂阿留申病毒（ADV）检测方法，根据GenBank中已登录的ADV基因组序列，选择水貂阿留申病毒VP2基因作为靶基因，设计并合成5套环介导等温扩增（LAMP）反应所需的引物，通过试验筛选出最佳引物，并进行最佳引物的特异性及敏感性试验，建立了LAMP快速检测方法。试验结果表明，该方法比普通PCR方法灵敏性高10倍，与其他的水貂源病毒不发生非特异性反应，并且具有良好的重复性。利用建立的LAMP检测方法对30份临床样品的阳性检出率为6.7%。该方法简便快速，为水貂阿留申病的检测提供了新的发展方向，有望成为简易的常规检测手段，尤其适用于基层应用。%The aim of the study was to develop a simple,rapid and specific assay for ADV detection. Based on the genomic sequence data of ADV in GenBank,five primers were designed and synthesized with VP2 as target gene. By comparing specificity and sensitivity of the primers,a LAMP assay was developed,which was proved to be 10 times more sensitive than normal PCR method and had no reaction with other mink viruses. The developed LAMP assay was used to test 30 suspected mink samples resulting in a positive rate of 6.7%. The assay was simple,rapid,reproducible and may be considered as a routine method used in grass-roots labs.