肾综合征出血热病毒76－118株的反转录及聚合酶链反应扩增、鉴定的初步研究

Reverse Transcription and cDNA Amplification of the 76-118 Strain of Hantann Virus

用反转录和聚合酶链反应（ＰＣＲ）技术扩增了肾综合征出血热病毒（ＨＦＲＳＶ）７６－１１８株ＭＲＮＡ３＇－端２３７ｂｐ基因片段。制备了长臂光敏氨基已酸生物素标记的２３７ｂｐ探针。经琼脂糖电泳和Ｓｏｕｔｈｅｒｎ印迹，生物素－亲和素系统杂交和显色，对２３７ｂｐ扩增产物进行了特异性和灵敏性鉴定。ＰＣＲ技术与反转录技术相结合可用来对一些ＲＮＡ病毒进行研究和诊断，并能快速制备出质量较好的标记探针。

Two-hundred and thirty-seven bp in M RNA from the 76-118 strain of Hantann virus was reverse transcribed and amplified by PCR.The long arm photobiotin-labelled specific 237bp probe was prepared.The results were shown in 2% agarose gel by EB and in Southern hybridization by photobiotin-Avidin system. The specificity and the sensitivity of the amplified products by PCR and Southern hybridization were characterized, less than 0. 1fg/ml concentration of target gene sequence could be detected by PCR and Southern hybridization. Results showed that the reverse transcription and PCR may also be used in other RNA viruses and the high sensitivity of nonradioisotopo-labelled probes such as photobiotin-Avidin system may be applied in the gene diagnoses and study.