傅万海 郝卫 彭宜君 510282 广州 第一军医大学附属珠江医院儿科 中华实验和临床病毒学杂志 1998 0 0 1


    关键词：轮状病毒；聚合酶链反应；地高辛素；斑点杂交 期刊 zhsyhlcbdxzz 0 病毒诊断 fur -->


    

【摘要】 目的 为探讨聚合酶链反应(PCR)技术标记的地高辛素(DIG)探针的特异性和敏感性。方法 用聚合酶链反应技术制备地高辛素标记的人类轮状病毒(HRV)cDNA探针，经cDNA-RNA斑点杂交。结果 该探针具HRV特异性，可检出10pg的RNA。应用该项技术检测了120份婴幼儿腹泻粪样标本，其阳性率为65%，明显高于PAGE方法(49.1%)的阳性率。结论 PCR方法直接制备地高辛素标记的cDNA探针方便、快速、特异性好、标记率高。

Detection of rotavirus RNA using DIG labelled probe prepared by polymerase chain reaction

Fu Wanhai, Hao Wei, Peng Yijun. Department of Pediatrics of Zhujiang Hospital. First Military University, Guangzhou 510282

Abstract The digoxigenin (DIG) labelled cDNA probe of rotaviruse was directly prepared by polymerase chain reaction(PCR). The results of cDNA-RNA hybridizaton showed that the DIG-cDNA probe exhibits rotavirus specificity and can detect as tiny as 10pg rotavirus RNA. 120 fecal samples of diarrhea from infants and young children were tested by dot-blot hybridization. It was shown that the positive rate of dot-blot hybridization(65.0%)was significantly higher than that of PAGE(49.1%). This study indicated that direct preparation of DIG-labelled rotavirus-cDNA probe by PCR is much faster and simpler than common method of labeling.

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