应用RT-nested-PCR检测犬唾液中狂犬病毒的初步研究

Preliminary study on detection of rabies virus from dog's saliva by RT-nested-PCR

目的建立RT-nested-PCR方法,用于检测可疑狂犬唾液中的狂犬病毒,为狂犬病的快速诊断提供一种新思路。方法根据N基因的保守区序列设计2对引物,通过反应条件的优化,建立RT-nested-PCR的方法,以扩增狂犬病毒的N基因,并用该方法检测可疑狂犬唾液中的狂犬病毒。结果该方法的最小RNA检出量可达11.2fg,并具有较好的特异性和重复性。结论建立的RT-nested-PCR检测法,特异性强,敏感度高,结果可靠,操作便捷,值得推广应用。

In the present study, a RT-nested-PCR assay was established for the detection of rabies virus in which two set of primers were designed from the nucleotide sequence of N gene,and the condition of RT-nested-PCR assay was validated. It was found that this assay could be used for the specific and stable detection of rabies virus from dog＇s saliva with a limit of detection of 11.2 fg RNA. It is concluded that a sensitive and rapid method for the detection of rabies virus from dog＇s saliva was successfully established, which provide a new means for further studies on the rapid diagnosis for rabies virus.