志贺菌耐多药相关基因水平转移的验证及其耐药性研究.pdfVIP

814 中国人兽共患病学报 2010 , 26 (9) Chinese Journal of Zoonoses 文章编号: 1002-2694(2010)09- 0814- 04 志贺菌耐多药相关基冈水平转移的 验证及其耐药性研究骨 王淑玲，宋春花，段广才，张卫东，都园林，未静後 摘 要:目的 对志贺菌耐多药相关基因TIC4) 水平转移进行验证.并对其与细菌耐药性的关系进行初步判定。方法 培养已筛选出的包含T1C4 基因片段的阳性克隆，提取其质粒.PCR 扩增 T1C4 基因片段、纯化、制备探针，与志贺菌敏感株 5523 、志贺菌耐多药转移株(5Tl l 人大肠埃希菌耐多药株(E667) 的基因组 DNA 和质粒 DNA 进行斑点杂交。采用药敏纸片 法研究含 T1C4 基因片段大肠埃希菌的耐药性。结果 应用 PCR 方法可以从 STl 1 和 E667 菌株基因组 DNA 中扩增出 T1C4 基因片段，而 5S23 菌株不含该基因片段;应用 T1C4 基因探针进行斑点杂交显示.STll 和 E667 菌株的全基因组和质粒 DNA 均可显示杂交信号，而 SS23 菌株的基因组和质粒 DNA 均不产生杂交信号; 14 种药物的药敏试验表明，含有TlC4 基因 的大肠埃希菌 (DH5α) 对四环素、先锋 V、头抱喔吩、氟吸酸、复方新诺明等五种药物的抑菌环直径明显小于(相差 3mm 以上) 不含 T1C4 基因的 DH5a 茵株。结论 TlC4 基因片段为 5Tll 菌株从 E667 菌株获得，且存在于两菌株的质粒上;TlC4 基因 可能介导细菌对多种药物的抗性。 关键词:志贺萄z 耐多药:基因水平转移 中图分类号:R378.2 文献标识码:A Identification on horizontal transfer of multi-drug resistance related gene fragment of Shigella and its drug-resistance WANG Shu-ling.SONG Chun-hua.DUAN Guang-cai.ZHANG Wei-dong.XI Yuan-lín.ZHU ling-yuan (Deþartmentof Eρidemiology. College of Puhlic Health. Zhengzhou University. Zhengzhou 450001. China) ABSTRACT: The objective of the present study was to identify horizontal transfer of gene fragment (T1C4) from E. coli to Shigella and its relationship with multi-drug resistance. Screened positive c10ne which contains T1C4 gene fragment was cul- tured and the plasmid was extracted. The fragment of T1C4 gene was then amplified by PCR and purified by gel purification kit. The probe was prepared and the dot hybridization was conducted for genomic DNA and plasmid DNA of Shigella sensitive strain (SS23). transferred multi-drug resistant Shigella strain (5Tll) and Escherichia colimulti-drug resistant cli