中国寄生虫学与寄生虫病杂志 ›› 1993, Vol. 11 ›› Issue (3): 172-175.

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多聚酶链反应分析溶组织内阿米巴的致病性

程训佳,Hiroshi Tachibana,Seiki Kobayashi,Yoshimasa Kaneda,黄美玉   

  1. 上海医科大学寄生虫学教研室; Tokai
  • 出版日期:1993-08-31 发布日期:2017-01-06
  • 基金资助:
    日本松前国际友好财团 Ohyama Health Foundation等的资助,在日本东海大学医学部感染症实验室完成,并受上海医科大学校基金

ANALYSIS OF PATHOGENICITY OF ENTAMOEBA HISTOLYTICA BY POLYMERASE CHAIN REACTION

  • Online:1993-08-31 Published:2017-01-06

摘要: 从急性阿米巴痢疾患者粪便中分离所得的溶组织内阿米巴虫株SH-3、SH-6、SH-8的DNA和包囊携带者粪便中分离所得的溶组织内阿米巴 SH-5、SH-7的 DNA应用多聚酶链反应技术扩增30个周期后作琼脂糖凝胶电泳分析,发现致病性引物 P_(11)、P_(12)可使SH-8虫株的DNA增殖;而非致病性引物P_(13)、P_(14)可使SH-3、SH-5、SH-6和SH-7虫株的DNA增殖。另外,酶株群分析和单克隆抗体反应也显示SH-8为致病性虫株,而另4株虫株均属于非致病性虫株,与多聚酶链反应的结果相一致。提示,致病性虫株与非致病性虫株的基因有区别,多聚酶链反应对溶组织内阿米巴进行基因分析是一种敏感和特异的新方法。

关键词: 溶组织内阿米巴, 多聚酶链反应, 引物, 酶株群, 单克隆抗体

Abstract: DNA from five isolates of Entamoeba histolytica were examined for their pathogenicity by polymerase chain reaction. Three isolates SH-3,SH-6,SH-8 were isolated from patients with acute amoebic dysentery, whereas SH-5 and SH-7 were isolated from asymptomatic cyst passers. Gel electrophoresis of PCR products showed that primers P11 , P12 for pathogenic strains could amplify genomic DNA extracted from SH-8 , and primers P13, P14 for non-pathogenic strains could amplify genomic DNA extracted from SH-3, SH-5, SH-6 and SH-7. Furthermore, zymodeme analysis and the reactivity of McAb 4G6, which recognizes the 30 kDa antigen of pathogenic E. histolytica indicated that only SH-8 was pathogenic, while the others were nonpathogenic. The results of the genotypic analysis by PCR were in accord with the phenotypic properties.It is suggested that there are differences in genomic DNA between pathogenic and non-pathogenic strains. PCR is a highly sensitive and specific method for genomic DNA analysis of E. histolytica.