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用限制性内切酶分析溶组织内阿米巴的致病性

PATHOGENICITY ANALYSIS OF ENTAMOEBA HISTOLYTICA AFTER RESTRICTION-ENDONUCLEASE TREATMENT

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【作者】 程训佳Hiroshi TachibanaSeiki KobayashiYoshimasa Kaneda黄美玉

【Author】 Cheng Xun-Jia, Hiroshi Tachibana, Seiki Kobayashi,Yoshimasa Kaneda, Huang Mei-Yu(Department of Parasitology, School of Basic Medical Sciences, Shanghai Medical University, Shanghai; Department of Infection Diseases, School of Medicine, Tokai University; Department of Tropical Medicine and Parasitology, School of Medicine, Keio University, Japan)

【机构】 上海医科大学基础医学院寄生虫学教研室日本国东海大学医学部感染症学教研室庆应大学医学部热带医学和寄生虫学教室上海医科大学基础医学院寄生虫学教研室

【摘要】 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。

【Abstract】 DNA from five strains of Entamoeba histolytica, SH-3, SH-5, SH-6, SH-7 and SH-8 were amplified 85 cycles by the polymerase chain reaction (FOB) followed by digestion by restriction-endonuclease Hinfl and EcoT22I resulted in -581-bp products. Hinfl digestion of SH-3, SH-5, SH-6 and SH-7 FOR products consistently yielded the same three fragments. The digestion pattern obtained was identical to that of the nonpathogenic SAW 142 control strain. The digestion pattern of SH-8 FOR products was identical to that of the pathogenic’ SAW 408 strain, and it DNA was similarly split into 2 fragments by EcoT22I. These observations indicate that 4 strains of E. Bistolytica isolated from cyst carriers and patients with fever and diarrhea but without purulent discharge are genotypically nonpathogenic, whereas SH-8 derived from patient with fever, diarrhea and purulent bloody-stool is genotypically pathogenic. These results completely correlate with zymodeme analysis. The genotypical identification of E. Histolytica using FOR and restriction-endonuclease digestion of amplified genomic DNA is Tery valuable.

【基金】 日本松前国际友好财团资助;Ohyamc Health Foundation资助;上海医科大学校基金资助
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