PCR制备地高辛素标记的探针检测轮状病毒核酸  

Detection of Nucleic Acid of HRV by a Dig-Labelled Probe Prepared with Polymerase Chain Reaction(PCR)

作　　者：卓礼梅[1] 徐帆[1] 陈火胜[1] 严泳楷 

机构地区：[1]中山医科大学微生物学教研室,中山大学昆虫所生物工程室

出　　处：《中国病毒学》1995年第1期22-27,共6页Virologica Sinica

摘　　要：用聚合酶链反应（ＰＣＲ）技术，制备了轮状病毒第９基因部分片段的地高辛标记的ｃＤＮＡ探针。ＤＮＡ－ＲＮＡ斑点杂交表明该探针具有轮状病毒Ａ组的特异性，可检出１０ｐｇＨＲＶ－ＲＮＡ。实验中选择了粪便标本提取核酸后点膜和粪便上清直接点膜进行斑点杂交，两种方法的结果一致；同时将斑点杂交法与ＰＡＧＥ法的结果进行了比较。实验结果表明用ＰＣＲ技术直接制备地高辛索标记的ｃＤＮＡ探针具有方便、快速、标记率高、特异性强的特点，具有一定的应用前景。cDNA probe of the partial genome segment of URV nineth gene was labelled with DIG by thetechnlque of PCR.DNA-RNA dot blot hybrid izat ion results showed the proporties of HRV A groupspeci ficity of this probe,The results were consistent by the method or loading the sam ple supornatantdirectly or loading after the extraction of fecal sample nucleic acld.We also compared the resul ts ofdot blot hybridlZation and PAG E.It was strongly ind icated in this ex periment that the Dig-labelledHRV cDNA probe prepared directly by PcR appeared to be convenient,fast,high labelling rate andhighly specific.

关 键 词：聚合酶链反应 地高辛素标记 斑点杂交 轮状病毒 

分 类 号：R392－33[医药卫生—免疫学] R373.24[医药卫生—基础医学]