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为呼吸道合胞病毒（ＲＳＶ）感染的临床诊断寻求敏感、特异的检测方法。采用ＲＴ－ＰＣＲ技术扩增病毒培养液和急性呼吸道感染患者鼻咽分泌物中的ＲＳＶ基因，并应用地高辛标记ｃＤＮＡ探针对扩增产物进行鉴定。结果成功地提取了ＲＳＶｍＲＮＡ，反转录合成ｃＤＮＡ，经扩增得到４７１ｂｐ左右ｃＤＮＡ区带。流感病毒Ｂ、副流感病毒２型、腺病毒７型对照无扩增带。ＲＳＶ培养液１０倍连续稀释至１∶１０００，经ＲＴ－ＰＣＲ扩增仍可见４７１ｂｐｃＤＮＡ区带。１３例患者鼻咽分泌物中的ＲＳＶ基因得到扩增，并经Ｓｏｕｔｈｅｒｎ印迹与斑点杂交证实。建立了ＲＳＶＲＴ－ＰＣＲ的方法，特异性、敏感性均较好，在临床标本的检测中将会有重要应用。
To find a sensitive and specific method for the clinical diagnosis of respiratory syncytial virus (RSV) infection. The RSV gene in nasopharyngeal secretions of patients with virus and culture of acute respiratory tract was amplified by RT-PCR, and the amplified product was identified by digoxigenin-labeled cDNA probe. Results The RSV mRNA was successfully extracted and cDNA was reverse transcribed. The cDNA fragment of 471 bp was amplified. Influenza virus B, parainfluenza virus type 2, adenovirus type 7 control without amplification band. RSV culture medium was diluted 10-fold to 1: 1000 and the 471 bp DNA band was still visible by RT-PCR. The RSV gene in the nasopharyngeal secretions of 13 patients was amplified and confirmed by Southern blotting and spot blotting. The established method of RSVRT-PCR, specificity, sensitivity are good, in the detection of clinical specimens will have important applications.