摘要
应用光敏生物素标记ｃＤＮＡ探针作打点杂交检测柯萨奇Ｂ＿３病毒（ＣＶＢ３）－ＲＮＡ，并采用薄层层析扫描仪反射扫描杂交信号和计算机整合反时峰面积的手段，对所测ＣＶＢ３－ＲＮＡ进行半定量。结果显示反射峰面积与进行自身杂交的ｃＤＮＡ含量（ｒ＝０．９８１，Ｐ＜０．００２５）及与病毒稀释度（ｒ＝０．９９７，Ｐ＜０．００５）呈有显著意义的直线正相关，提了薄层层析扫描结合核酸杂交的确可对ＣＶＢ３－ＲＮＡ进行半定量分析，也可为研究其他病毒或非病毒核酸提供新的检测方法，值得推广。
etection of Coxsackie B-3 virus RNA was done by dot-blot hybridisation using enterovirus RNA specific cDNA probe(C B2-cDNA) labelled with photobiotin,and half quantitation of hybridisation signals was investigated by scanning densimetry and computerizing integration of reflec-tion peaks'area.The result showed that the lineal relation between areas of reflection peaks and the known amount of purified CB2-cDNA(r=0,981,P< 0.0025) or the dilution of Coxsackie B-3 virus stock suspension(r=0.997,P<0.005) was significant.It suggested that Coxsackie B-3 virus RNA could be half-quantitated by the above mentioned method.In addition,the photobiotin-labelled CB2-cDNA probe was highly sensitive and specific, and the technique of labelling was very simple.It may serve as a general detective method and half quantitative analysis of Coxsackie B virus RNA and other viral genome DNA or RNA.
基金
上海市科委资助课题
