论文部分内容阅读
目的建立快速筛选细菌耐药整合子的分类方法,并对21株来自临床的菌株进行整合子筛选和分类。方法根据G enB ank/EM BL内的第一、第二和第三类的整合酶序列,通过软件C lustalW的多重比对分析设计出各类整合子的特异性引物,用对照菌株建立多重PCR方法并对21株临床菌株进行PCR扩增,通过其PCR扩增产物片段大小的不同进行整合子分类。结果对照菌株实验结果显示,21株临床分离菌株中,15株在565bp处有扩增片段,即为含有第一类整合酶基因;4株在403bp处有扩增片段,即含有第二类整合酶基因;1株在565bp和403bp处均有扩增片段,提示其同时含有第一、二类整合酶基因;1株没有得到任何扩增片段,即不含有这三类整合酶基因;在被测菌株中未发现第三类整合酶基因的阳性菌株。结论本文报道的筛选细菌耐药整合酶基因分类的多重PCR方法效果良好,具有可行性,为更加全面细致研究整合子类型以及整合子介导的细菌耐药机制提供了一种简单易行、快捷有效的方法。
OBJECTIVE: To establish a rapid screening method for the classification of bacterial resistance integron, and to screen and classify 21 strains of clinical isolates. Methods Based on the first, second and third integrase sequences in G enB ank / EM BL, specific primers of various integrons were designed by multiple alignment analysis of software C lustal W. Multiplex PCR Methods 21 strains of clinical isolates were amplified by PCR and subcloned according to the size of their PCR products. Results The results of the control strains showed that among the 21 clinical isolates, 15 were amplified at 565bp, which contained the first type of integrase gene. The 4 strains contained an amplification fragment at 403bp, which contained the second type of integration 1 gene at 565bp and 403bp both amplified fragment, suggesting that it contains both the first and second class integrase gene; 1 did not get any amplified fragment, that does not contain these three types of integrase gene; was No positive strains of the third type of integrase gene were found in the tested strains. Conclusion The multiplex PCR method reported in this paper for screening bacterial resistance to integrase gene is effective and feasible. It provides a simple, quick and easy method for more comprehensive and meticulous research on the integration subtype and integration-mediated mechanism of bacterial resistance effective method.