【摘要】 目的　建立一种筛选第一、二和三类整合酶基因 (intⅠ )的简并引物PCR方法 ,并对临床菌株进行整合子筛选和分类。方法　用梯度PCR方法确定PCR反应的最适退火温度 ,并对 1 6株临床标本进行PCR扩增 ,通过对阳性PCR扩增产物的限制性内切酶分析进行整合子分类。结果　根据梯度PCR结果确定最适退火温度为 4 6℃。应用简并引物PCR法检测 1 6株临床分离株 ,其中 8株阳性 ,经酶切分类后确定其中 4株只含有第一类整合酶基因 ,有 3株含有第二类整合酶基因 ,1株同时含有第一类和第二类整合酶基因。结论　建立了一种同时筛选第一、二和三类整合子的方法 ,实际检测显示其可行性 ,为全面细致研究整合子介导的细菌耐药提供了一种快速、简便的方法。更多还原

【Abstract】 Objective To construct a PCR assay using degenerate primers to screen class 1,2 and 3 integrase (intⅠ). Methods Degenerate primers were designed and then gradient PCR was used to decide the optimization annealing temperature. The PCR amplicons were analyzed by digestion with restriction endonucleases. Results 46℃ of optimization annealing temperature was decided according to the gradient PCR results. 16 of clinical strains were used in this study, 8 of strains were positive for PCR. The amplicons digested with restriction endonucleases showed that 4 of strains harbored intⅠ1, 3 of strains harbored intⅠ2, 1 of strains harbored both intⅠ1 and intⅠ2. Conclusion A PCR assay to screen class 1, 2 and 3 integron at one time was established and confirmed to be practicable by application. A swift and versatile method is provided for comprehensive and particular research of antibiotic resistance genes carried by integrons in bacteria.更多还原