Object: To establish reverse transcription nested-polymerase chain reaction(RT-n-PCR)with universal primers, in order to detect enterovirus distinctively and rapidly. Methods: RT-n-PCR was established with two universal primers within 5' untranslated region(5'-UTR) of enterovirus. Three standard enteroviruses, three standard non-enteroviruses and 327 clinical specimens obtained from 165 patients suspected enterovirus infection were detected with RT-n-PCR. Results: Clear bands were seen at 312 base pair position of gel electrophoresis in the 3standard enteroviruses, while none was seen in the 3 standard non-enteroviruses. Bands were also seen at the same position in 104 clinical specimens. The enterovirus positive rate of clinic specimen was 31.8%. Both stool and throat swab were detected in 147 patients. 49 stools and 44 throats swab specimens were positive respectively. 35 specimens of them were positive both in stool and throat swab. There was no significant difference of enterovirus positive rate between stool and throat swab (P＞0.05). Conclusions: RT-n-PCR could detect enterovirus accurately with high specificity and little time, it could raise the enterovirus positive rate by detecting various specimens.%目的设计通用引物建立巢式逆转录聚合酶链反应(RT-n-PCR),以达到广泛.特异及快速诊断肠道病毒(Enterovirus,EV)感染.方法在EV基因组5'端非编码区(5'-UTR)设计二对通用第物,建立RT-n-PCR检测方法,对3种EV和3种非EV标准株以及165例可疑EV感染的327份临床标本进行检测.结果EV标准株均在电泳中检测到一清晰312bp扩增条带,而非EV标准株则无扩增条带;共有104份标本检测到相应大小扩增条带,阳性率31.8%;147例病例同时检测了粪便和咽拭子,粪便阳性49例,咽拭子阳性44例,其中粪便和咽拭子同时阳性35例,粪便和咽拭子EV阳性比较无显著性差异(P＞0.05).病例总阳性数64例,阳性率38.8%.结论RT-n-PCR可以准确地检测出肠道病毒,特异性强,用时少,同时检测多份标本可以提高EV阳性率.
展开▼
