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PCR制备地高辛素标记的探针检测轮状病毒核酸
引用本文:卓礼梅,徐帆,陈火胜,严泳楷. PCR制备地高辛素标记的探针检测轮状病毒核酸[J]. Virologica Sinica, 1995, 0(1)
作者姓名:卓礼梅  徐帆  陈火胜  严泳楷
作者单位: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
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。

关 键 词:聚合酶链反应(PCR),地高辛素标记,斑点杂交,轮状病毒(HRV)

Detection of Nucleic Acid of HRV by a Dig-Labelled Probe Prepared with Polymerase Chain Reaction(PCR)
Zhuo Limei,Xu Fan,Chen Huosheng,Yan Yoiigkai. Detection of Nucleic Acid of HRV by a Dig-Labelled Probe Prepared with Polymerase Chain Reaction(PCR)[J]. 中国病毒学(英文版), 1995, 0(1)
Authors:Zhuo Limei  Xu Fan  Chen Huosheng  Yan Yoiigkai
Abstract: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.
Keywords:Polymerase chain reaction(PCR)  DIG-labelling  Dot blot hybridization  Human ro tavirus(HRV)  
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