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用SH-3、SH-5、SH-6、SH-7和SH-85株溶组织内阿米巴的DNA增殖35个周期,将其基因DNA用内切酶HinfⅠ和EcoT22Ⅰ进行消化后,作琼脂糖凝胶电泳分析,显示,5株虫株DNA经35个循环周期后产生531-bp的产物;经HinfⅠ消化后,SH-3、SH-5、SH-6和SH-7的基因DNA产生3个相同的片段,而显示其均与非致病性虫株SAW142的电泳谐完全相同;而SH-8基因DNA的电泳谱与致病性虫株SAW408的电泳谱一致,而用EcoT22Ⅰ消化结果也显示SH-8的图谱与致病性虫株SAW408的相同,证实了从包囊携带者和有发热、腹泻而无脓血便患者粪便中分离的虫株均属于非致病性虫株,从有发热、腹泻、脓血便的急性阿米巴痢疾患者粪便中分离到的虫株属于致病性虫株,均与酶株群分析相一致。提示,应用多聚酶链反应和限制性内切酶消化基因DNA来检测溶组织内阿米巴的基因型是十分有意义的。
The DNA of Entamoeba histolytica was propagated for 35 cycles with SH-3, SH-5, SH-6, SH-7 and SH-85 strains, digested with HinfⅠ and EcoT22Ⅰ, Agarose gel electrophoresis analysis showed that the DNA of the 5 strains produced 531-bp products after 35 cycles. After HinfⅠ digestion, the DNA of SH-3, SH-5, SH-6 and SH-7 3 identical fragments were obtained and showed the same electrophoretic resonance as the non-pathogenic strain SAW142. The electrophoresis pattern of SH-8 gene DNA was consistent with that of the pathogenic strain SAW408, and digested with EcoT22I The results also showed that the pattern of SH-8 was the same as that of the pathogenic strain SAW408 and confirmed that the isolates belonging to the non-pathogenic strain were isolated from the encapsulated carriers and the feces from patients with fever and diarrhea without sepsis, Insect strains isolated from the feces of patients with fever, diarrhea and pus and blood from patients with acute amebiasis are pathogenic and are consistent with enzyme group analysis. It is of great significance to detect the genotype of Entamoeba histolytica using polymerase chain reaction and restriction endonuclease digestion of genomic DNA.