取定量的HK-9株溶组织内阿米巴滋养体放入受检者的0.1g粪便中,再加DTT、KOH等在65℃孵育15min,后用HCl中和,并以酶/氯仿/异戊醇振荡抽提DNA。同时再取受检者粪便0.1g以NaCl洗涤,离心,蛋白酶K消化,抽提其DNA。用以rRNA为模板设计的致病性与非致病性引物进行双向PCR,PCR产物以含0.2μg/ml溴乙锭的1.3％的琼脂糖凝胶电泳显示,并用限制性内切酶DraⅠ和Sau 96Ⅰ进行消化。 The stained HK-9 strains of Entamoeba histolytica trophozoites were placed in 0.1 g of the stools of the subjects, incubated with DTT, KOH and the like at 65 ° C for 15 min, neutralized with HCl and treated with enzyme / chloroform / Isoamyl alcohol is shaken to extract DNA. At the same time take the test manure 0.1g was washed with NaCl, centrifuged, digested with proteinase K, extracted DNA. Two-way PCR was performed with pathogenic and non-pathogenic primers designed using rRNA as a template. The PCR product was displayed on a 1.3% agarose gel containing 0.2 μg / ml ethidium bromide and was digested with restriction endonucleases Dra I and Sau 96 I for digestion.