水貂肺炎克雷伯菌的分离鉴定及耐药性分析

doi:10.16431/j.cnki.1671-7236.2022.02.032

摘要/Abstract

摘要： 【目的】肺炎克雷伯菌是人畜共患的条件致病菌, 是引起水貂肺炎的常见病原菌之一。试验主要对吉林省某水貂养殖场送检的6只患有肺炎的病死水貂进行细菌分离鉴定及耐药性分析, 为指导临床合理用药提供依据。【方法】通过分离纯化方法从样品中分离菌株并进行PCR鉴定, 应用多位点序列分型技术(MLST)进行菌株ST分型。通过感染小鼠了解菌株的致病性, PCR检测血清型及毒力基因的携带情况; 运用琼脂稀释法检测分离菌株对18种药物的敏感性, 并通过PCR检测分离菌株耐药基因的携带情况。【结果】分离得到的6株菌株经PCR鉴定为肺炎克雷伯菌; MLST分析结果表明6株菌株分别有6种ST型, 分别为: ST2594、ST1782、ST2844、ST2330、ST86和ST661。致病性分析结果显示, 6株菌株均可引起小鼠死亡, 共检测出6种毒力基因, 未检测到毒力较强的血清型。药敏试验结果显示, 6株肺炎克雷伯菌对头孢他啶、头孢曲松、亚胺培南、美罗培南、阿米卡星较敏感, 对链霉素、庆大霉素、卡那霉素、青霉素、氨苄西林、环丙沙星、恩诺沙星、左氧氟沙星、多西环素和氟苯尼考较耐药。耐药基因检测结果显示, 6株菌株共检测出bla_SHV、bla_TEM-1、bla_CTX-M-1G、aadA1、aac(3’)-Ⅳ、aac(3’)-Ⅱc、aph(4’)-Ⅰa、aph(3’)-Ⅶ、aph(3’)-Ⅳ、aph(2’)-Ⅰb、rmtB、qnrB、qnrD、qnrS、qepA、oqxAB、floR和mcr-1 18种耐药基因。【结论】分离到的6株肺炎克雷伯菌株为不同的种系进化, 并具有较强的致病性和耐药性, 为临床用药治疗带来困难。

关键词: 水貂; 肺炎克雷伯菌; 药物敏感性; 耐药基因

Abstract: 【Objective】 Klebsiella pneumoniae is a zoonotic opportunistic pathogen, and also one of the common pathogenic bacteria causing mink pneumonia.In order to guide rational drug use and provide the basis for treatment to 6 dead minks suffering from pneumonia provided by mink farms in Jilin province, the bacterial isolation and identification, drug resistance spectrum and drug resistance gene analysis were carried out.【Method】 Strains were isolated from samples by isolation and purification method, and identified by PCR method, and ST typing was performed by multilocus sequence typing technique(MLST) method.The pathogenicity of the strain was learned by infecting mice.Serotype and the virulence gene was detected by PCR.The susceptibility of the strains to 18 drugs was detected by agar dilution method, and the carrying status of drug-resistant genes was detected by PCR.【Result】 The results showed that 6 isolates were identified as Klebsiella pneumoniae by PCR.MLST analysis results showed that 6 strains had 6 ST types, namely ST2594, ST1782, ST2844, ST2330, ST86 and ST661, respectively.Pathogenicity analysis results showed that all 6 strains could cause the death of mice, and a total of 6 virulence genes were detected in the isolates.No more virulent serotypes were detected.The results of drug sensitivity test showed that 6 strains of Klebsiella pneumoniae were more sensitive to ceftazidime, ceftriaxone, imipenem, meropenemand amikacin, and more resistant to streptomycin, gentamicin, kanamycin, penicillin, ampicillin, ciprofloxacin, enrofloxacin, levofloxacin, doxycycline and florfenicol.There were 18 resistance genes detected in 6 strains, including bla_SHV, bla_TEM-1, bla_CTX-M-1G, aadA1, aac(3')-Ⅳ, aac(3')-Ⅱc, aph(4')-Ⅰa, aph(3')-Ⅶ, aph(3')-Ⅳ, aph(2')-Ⅰb, rmtB, qnrB, qnrD, qnrS, qepA, oqxAB, floR and mcr-1.【Conclusion】 6 isolates of Klebsiella pneumoniae had different phylogenetic evolutionand strong pathogenicity and drug resistance, they brought difficulties to clinical drug treatment.

Key words: mink; Klebsiella pneumoniae; drug sensitivity; resistance gene

中图分类号:

S852.61⁺2

陈强, 程悦宁, 冯秋菊, 郭利, 张淑琴, 谭斌, 易立, 赵权, 程世鹏, 孙娜. 水貂肺炎克雷伯菌的分离鉴定及耐药性分析[J]. 中国畜牧兽医, 2022, 49(2): 700-708.

CHEN Qiang, CHENG Yuening, FENG Qiuju, GUO Li, ZHANG Shuqin, TAN Bin, YI Li, ZHAO Quan, CHENG Shipeng, SUN Na. Isolation, Identification and Drug Resistance Analysis of Klebsiella pneumoniae from Minks[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 700-708.

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