摘要目的 利用聚合酶链式反应(PCR)技术,建立鼠疫耶尔森菌(简称鼠疫菌)多种耐药基因的检测方法,为鼠疫的治疗提供指导性建议.方法 根据美国国立生物信息中心(NCBI)公布的耐氨基糖苷类链霉素strA、strB基因、耐β-内酰胺类抗菌药物tem、shv、ctx-m基因,耐磺胺类药物sul1、sul2、sul3基因序列,分别对每个基因设计1对引物,对282株分离自青海省鼠疫自然疫源地鼠疫菌的DNA进行PCR扩增,扩增产物进行凝胶电泳,用凝胶成像系统拍照后读取结果.对282株菌株进行链霉素、磺胺甲恶唑、头孢曲松钠3种药物的耐药性检测.结果282株菌株的PCR扩增结果均为阴性,尚未发现具有链霉素、磺胺类药及β-内酰胺类抗菌药抗药基因的菌株.药敏试验显示,282株菌株对链霉素、磺胺甲恶唑、头孢曲松钠均高度敏感.抑菌环直径分别 >19、17、21 mm.结论 利用PCR技术作为鼠疫菌多种耐药基因的检测方法是可行的,应用该方法能系统监测大批量鼠疫菌的耐药基因是高效、经济、实用的实验方法,能为鼠疫治疗提供指导性建议.

abstractsObjective To establishment a method for detection of multiple drug resistance gene of Yersina pestis using polymerase chain reaction(PCR), to provide a guidance for treatment of plague. Methods According to National Center for Biotechnology Information (NCBI) released sequences of aminoglycoside resistant genes of streptomycin resistant,strB,strA,beta lactam antibiotics resistant genes tem,shv,and ctx-m,sulfamilamide resistant genes sul1, sul2, and sul3, a pair of primers of each gene was designed. DNAs of 282 strains isolated from plague natural foci in Qinghai Province were amplified by PCR using every pair of primers. The products were separated using gel electrophoresis, and the results were visualized through a gel imaging system. The susceptibility of 282 Yersina pestis to streptomycin, sulfamethoxazole and ceftriaxone was tested by drug sensitivity test. Results The PCR amplification results of all samples were negative,and strains with streptomycin,sulfamilamide and beta lactam antimicrobial drug resistance genes were not found. Drug sensitivity test showed that 282 strains were highly sensitive to streptomycin,sulfamethoxazole and ceftriaxone sodium.The diameter of bacteriostasis ring>19,17,21 mm, respectively. Conclusions It is a feasible method to use PCR technology to detect the multiple drug resistance genes of Yersinia pestis. Using this method to systematically monitor the resistance gene of Yersinia pestis is an efficient, economical and practical experimental method, which can provide guidance for the treatment of plague disease.