Nested PCR检测狂犬病病毒方法的建立及病毒在小鼠体内的移行动态

Detection of Rabies Virus by Nested PCR and Spread of Virus to Peripheral Tissues in Mice

应用建立的 Nested PCR特异地检出狂犬病病毒株 CVS、 HEP-Flury、ERA、RC-HL、 1 0 88、Komatsugawa的 RNA,但对类狂犬病病毒 Lagos bat、Duvenhage、Mokola及水泡性口膜炎病毒、轮状病毒、犬瘟热病毒均为阴性。该法敏感性很高 ,能检出 3TCID5 0 或 0 .8pg的狂犬病病毒 RNA。用该法测定了小鼠脑内感染 CVS株后的病毒增殖和移行动态 ,对感染小鼠的主要内脏器官进行了病毒 RNA检测 ,结果发现小鼠脑内感染 CVS 5d以后在其心、肝、脾、肺等内脏器官均检出了病毒 RNA。

A nested PCR has been successfully developed to detect rabies virus. It could detect a volume of 3 TCID 50 of rabies virus and gave a positive result of 0.8 pg of RNA. The positive results were only from rabies virus strains CVS, HEP flury, RC HL, ERA, 1088, Komatsugawa by two pairs of primers, while the rabies related virus strains Lagos bat, Duvenhage, Mokola and other virus strains VSV, rotavirus and CDV showed negative. In the detection of growth of rabies virus in mice brain, Nested PCR provided a positive result in the first two days after inoculation, however IFA and PCR showed negative. Nested PCR could also identify RNA of rabies virus in liver, heart, lung, spleen of mice 5 days after inoculation.