用RT-PCR技术检测石蜡包埋组织中的肠道病毒RNA

Detection of the enterovirus RNA by RT-PCR from paraffin-emmbedded tissue

目的 :应用逆转录 PCR技术 ( RT- PCR)在石蜡包埋的心肌组织切片中检测肠道病毒 (柯萨奇病毒 B3 ,CVB3 ) RNA。方法 :用 Trizol法从石蜡包埋的心肌组织切片中提取 CVB3 RNA,用适当的引物合成 c DNA第一链 ,从中取出适量进行聚合酶链反应 ( PCR)。PCR结果用琼脂糖凝胶电泳检测并将 PCR产物做核苷酸序列分析。阳性对照采用 CVB3 感染的人羊膜细胞。结果 :用RT- PCR技术可扩增出一条约 5 0 0 bp( 5 4 0 bp)的核苷酸片段 ,测序结果表明是 CVB3 的一个片段。结论 :RT- PCR技术应用于长期保存的石蜡包埋的组织切片进行肠道病毒的检测可获得较满意结果 ,可将 RT-

Objective:To detect enteroviruses RNA in paraffin embedded tissue by RT PCR. Methods:To synthesis cDNA using total RNA extracted from the paraffin embedded tissue, then using a pair of propriate primes to produce considerable DNA.The products of PCR were detected by agarose gel electrophresis and sequenced as well. The positive control were culture cells infected by CVB 3. Results: The product of RT PCR was passage of 540bp and its sequence was identical to that of CVB 3. Conclusion: RT PCR is a practicable method to detect enterovirus RNA in paraffin embedded tissue and it will be a good method for reviewing research.