广东省羊源肺炎克雷伯菌遗传进化与毒力基因及耐药性分析

旨在了解广东省羊源肺炎克雷伯菌毒力基因、耐药基因、耐药谱构成以及遗传多样性。本研究从广东省3个市的不同规模化羊场采集了150份羊鼻拭子样品,通过分离纯化、染色镜检等方式进行致病菌的分离鉴定,利用分子生物学以及依据CLSI手册的药敏试验等方法进行分离菌株的毒力基因、耐药性等生物学特性研究。结果显示：从样品中分离得到42株肺炎克雷伯菌,药敏试验结果显示,阿莫西林以及氧氟沙星耐药率高达100%,对其余药物均有不同程度耐药性;毒力基因检测结果显示,rmpA、kfuBC、ureA分别为88.0%、85.7%和83.3%;耐药基因检测结果显示,耐药基因aphA、qnrA、aacC4、aacC2的检出率为42.9%、40.5%、28.6%、23.8%,其余耐药基因检出率较低。ERIC-PCR分型结果显示,相似性>80%的可分为7个类群,其中,群体Ⅰ为优势群体,占比66.6%。本试验对广东地区羊源肺炎克雷伯菌的毒力基因、流行状况和耐药性进行分析研究,对羊源肺炎克雷伯菌的防治有一定的指导意义。

Isolation,Identification and Biological Characteristics of Klebsiella pneumoniae from Sheep in Guangdong Province

This study investigated the virulence genes, drug resistance genes, drug resistance spectrum and genetic diversity of Klebsiella pneumoniae from sheep in Guangdong province. One hundred and fifty sheep nose swab samples were collected from sheep farms with different scales in 3 cities of Guangdong province. The pathogenic bacteria were isolated and identified by means of culture, purification, staining and microscopic examination. The biological characteristics of the isolated strains, such as virulence genes and drug resistance etc., were studied by means of molecular biology and drug sensitivity test according to CLSI manual. The results showed that 42 strains of K. pneumoniae were isolated from the samples. The drug susceptibility test results showed that the drug resistance rate of amoxicillin and ofloxacin was up to 100%, and there were different degrees of resistance to other drugs; the virulence gene test results showed that rmpA, kfuBC and ureA were 88.0%, 85.7% and 83.3%, respectively. The detection results of drug resistance genes showed that the detection rates of drug resistance genes aphA, qnrA, aacC4, aacC2 were 42.9%, 40.5%, 28.6%, 23.8%, and the detection rates of other drug resistance genes were low. Eric-pcr typing results showed that the homology was more than 80.0%, which could be divided into seven groups, among which population Ⅰ was the dominant population, accounting for 66.6%.This study was conducted to analyze the virulence genes, epidemic status and drug resistance of sheep K. pneumoniae in Guangdong area, which has certain guiding significance for the prevention and control of sheep K. pneumoniae.