狂犬病病毒LAMP检测方法的建立及初步应用

Establishment and application of loop-mediated isothermal amplification for rabies virus

针对狂犬病病毒基因Ⅰ型核蛋白保守区设计4条识别靶序列上6个位点的特异性引物,在链置换聚合酶(Bst酶)作用下,60℃恒温1 h内完成扩增。反应特异性强、灵敏度高。扩增产物可通过琼脂糖凝胶电泳、显色反应或浊度比较进行判定。通过对反应条件的优化,病毒基因的的最低检出量可达到101拷贝数。该方法可用于狂犬病病毒的实验室检测和临床初步诊断。

The loop-mediated isothermal amplification（LAMP）was established which can detect rabies virus.Design specific primer set to identify six fragments of rabies virus neucleoprotein sequence.The reaction of LAMP was completed within 1h by Bst polymerase catalysis under isothermal condition.It could be detect 101copys of rabies virus.The specificity of LAMP products could be easily confirmed by agarose gel electrophoresis,color reaction or turbidity comparison.This method is ideal for detect rabies virus in laboratory and primary diagnosis of clinical infection.