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目的探讨动物食品中肺炎克雷伯菌质粒介导产Amp C酶的耐药表型与基因型,以控制动物食品中的细菌耐药性在动物、环境和人类之间的传播。方法采用头孢西丁纸片扩散法筛选产Amp C酶菌株;采用三维试验确证产Amp C酶;采用聚合酶链反应(PCR)和DNA测序检测分析Amp C酶基因型别;采用UNIQ-10柱式质粒小量抽提试剂盒提取细菌质粒。结果 67株肺炎克雷伯菌经头孢西丁敏感试验选出疑产Amp C酶菌株15株,经三维试验确证产Amp C酶菌株有12株,15株疑产Amp C酶菌株经PCR检测和DNA测序证实有10株菌株均为DHA-1型Amp C酶。结论动物食品中分离的肺炎克雷伯菌产Amp C酶的菌株检出率较高,基因型以DHA-1型为主,并且产Amp C酶的肺炎克雷伯菌表型与基因型不完全相同,二者的结果有一定的关联性但同时存在差异。临床兽医应慎用β-内酰胺类药物,动物食品加工更应严格操作规程,切断动物食品链传递给人类。
Objective To investigate the resistant phenotype and genotype of Amp C enzyme mediated by Klebsiella pneumoniae in animal food to control the spread of bacterial resistance in animal food between animals, environment and human beings. Methods AmpC enzyme was screened by cefoxitin disc diffusion method. AmpC enzyme was confirmed by three-dimensional test. Genotypes of AmpC enzyme were detected by polymerase chain reaction (PCR) and DNA sequencing. UNIQ-10 column Plasmid mini-extraction kit to extract bacterial plasmids. Results 67 strains of Klebsiella pneumoniae were selected by Ampicillin susceptibility test, and 15 strains of AmpC enzyme were confirmed by three-dimensional test. Fifteen strains of AmpC enzyme were detected by PCR and DNA sequencing confirmed that 10 strains were all DHA-1 type Amp C enzyme. Conclusions The detection rate of Amp C enzyme produced by Klebsiella pneumoniae in animal food is high, the genotype is DHA-1 type, and the phenotype and genotype of Klebsiella pneumoniae producing Amp C enzyme are not Exactly the same, the results of the two have some relevance but at the same time there are differences. Clinical veterinary drugs should be used with caution β-lactams, animal food processing should be more strict rules, cut off the animal food chain passed to humans.