小鼠诺如病毒巢式PCR检测方法的建立及应用

Development and Application of the Nested PCR Assay for Murine Norovirus

为了建立一种高敏感性和特异性的检测方法,突破目前小鼠诺如病毒(MNV)检测局限,根据MNV保守区基因组序列设计合成内、外2对引物,通过PCR反应条件的优化,建立MNV的巢式PCR检测方法。结果表明,该方法仅能特异的扩增MNV目的基因而不能扩增其他病毒,且敏感度达到500个拷贝,比普通PCR提高1000倍。说明本研究建立的巢式PCR方法具有特异性强、敏感性高、重复性好等优点,可以准确快速的用于小鼠诺如病毒的病原检测。

In order to seek a high sensitive and specific detection method to break through the MNV detection limits,we established a nested PCR assay for detecting MNV.Two pairs of primers were designed according to the conserved region of MNV genome sequences.The results showed that the nested PCR was 1000 times more sensitive than the conventional PCR,and it was only able to amplify the murine norovirus with the sensibility of 500 copies/μL.The results indicate that the established nested PCR method has high sensitivity and specificity,which can be used for a quick diagnosis and epidemiological investigation of MNV.