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多聚酶链反应是体外DNA增殖的新技术。在多聚酶、dNTP和引物的参与下在数小时内使特异性的DNA顺序大量增殖。用B、C、G、T4株溶组织内阿米巴的DNA增殖30周期后,作凝胶电泳分析,发现致病性虫株的引物不能使B、C、G、T虫株的DNA增殖,凝胶电泳不出现条带;而非致病性虫株的引物能使B、C、G、T虫株的DNA增殖,凝胶电泳出现特异性条带。用辣根过氧化物酶标记致病性虫株DNA探针与B、C、G、T的DNA进行斑点杂交呈阴性反应;而用非致病性虫株DNA探针与B、C、G、T的DNA斑点杂交呈阳性反应。实验结果显示B、C、G、T为非致病性虫株,与同工酶分析一致。多聚酶链反应是鉴别致病性和非致病性溶组织内阿米巴虫株的高特异性和敏感性方法,为从分子生物学研究溶组织内阿米巴的致病机理提供一新技术。
Polymerase chain reaction is a new technology in vitro DNA proliferation. Specific DNA sequences proliferate in a matter of hours with the aid of polymerase, dNTPs and primers. After 30 cycles of DNA amplification of Entamoeba histolytica strains B, C, G and T4, the results of gel electrophoresis analysis showed that the primers of the pathogenic strains could not proliferate the DNA of the strains B, C, G and T , Gel electrophoresis showed no bands; while non-pathogenic strains of primers can make B, C, G, T strain DNA proliferation, gel electrophoresis showed specific bands. The horseradish peroxidase-labeled DNA probe was used for spot-blot hybridization with the DNA of B, C, G and T, while DNA probe of non-pathogenic strain was used to detect the B, C, G , T DNA spot hybridization was positive. The experimental results show that B, C, G, T non-pathogenic strains, consistent with isozyme analysis. Polymerase chain reaction is a highly specific and sensitive method for the identification of pathogenic and non-pathogenic E. histolytica strains, providing a new technique for studying the pathogenesis of Entamoeba histolytica from molecular biology .