甘宁黄土高原阿拉善黄鼠鼠疫疫源地鼠疫菌携带耐药及耐消毒剂基因的调查

Investigation on antibiotic-resistance and disinfectant-resistant genes of Yersinia pestis carriers of Spermophilus dauricus alashanicus isolated in natural plague foci of Ganning loess plateau

目的了解甘宁黄土高原阿拉善黄鼠鼠疫疫源地内分离的鼠疫菌携带耐消毒剂及耐药相关基因的状况,为指导该疫源地在今后的鼠疫防治工作中合理使用抗菌药物和消毒剂提供参考依据。方法利用常规的十二烷基硫酸钠(SDS)裂解,酚-氯仿抽提方法对甘宁黄土高原阿拉善黄鼠鼠疫疫源地境内分离的49株鼠疫菌进行DNA染色体的提取,DNA终浓度稀释至0.002μg/μl,并对耐氨基糖苷类链霉素基因(strA、strB)、耐β-内酰胺类药物相关基因(tem、ctx-m),耐磺胺类药物相关基因(sul1、sul2)及耐季铵盐类消毒剂基因(qacEΔ1-sul1)7对引物按设定的PCR反应体系逐一进行PCR扩增,同时设阳性对照和空白对照。结果通过PCR扩增在49株鼠疫菌中未检测出耐氨基糖苷类链霉素基因(strA、strB)、耐β-内酰胺类药物相关基因(tem、ctx-m),耐磺胺类药物相关基因(sul1、sul2)及耐季铵盐类消毒剂基因(qacEΔ1-sul1)。结论甘宁黄土高原阿拉善黄鼠鼠疫疫源地分离的49株鼠疫菌不具有耐消毒剂及耐药相关基因的特征。

Objective To understand the status quo of antibiotic-resistance and disinfectant-resistant related genes of Yersinia pestis carriers of Spermophilus dauricus alashanicus isolated in natural plague foci of Ganning loess plateau, and to provide references and basis for rational use of antimicrobial agents and disinfectants during prevention and control in the foci in the future.Methods DNA chromosomes of 49 strains isolated from Yersinia pestis of Spermophilus dauricus alashanicus in the plague foci of Gansu loess plateau were extracted by conventional sodium dodecyl sulfate(SDS) cracking and Phenol-chloroform extraction method.The final concentration of DNA was diluted to 0.002 μg/μl, and seven pairs of primers of aminoglycoside-resistant streptomycin gene(strA,strB),β-resistant lactam agent related genes(tem,ctx-m),sulfonamide-resistant agent related genes(sul1,sul2) and quaternary ammonium salt-resistant disinfectant genes(qacEΔ1-sul1) were amplified by PCR according to PCR reaction system, and positive control and blank control were simultaneously set.Results Aminoglycoside-resistant streptomycin gene(strA,strB),β-resistant lactam agent related genes(tem,ctx-m),sulfonamide-resistant agent related genes(sul1,sul2) and quaternary ammonium salt-resistant disinfectant genes(qacEΔ1-sul1) were not detected in 49 strains of Yersinia pestis via PCR amplification.Conclusion Forty nine strains of Yersinia pestis of Spermophilus dauricus alashanicus isolated in plague natural foci of Ganning loess plateau aren’t characterized with disinfectant-resistant and antibiotic-resistance related genes.