分离自屠宰场的肺炎克雷伯菌的耐药性分析

doi:10.11843/j.issn.0366-6964.2023.07.035

摘要/Abstract

摘要： 为了解猪源肺炎克雷伯菌（Klebsiella pneumoniae）分离株的耐药情况和毒力基因携带情况，本试验对广州市某屠宰场进行采样调查和耐药性分析。通过对猪肺样本进行肺炎克雷伯菌分离鉴定，对分离株进行6种常见的荚膜血清型（K1、K2、K5、K20、K54、K57）鉴定，并用琼脂扩散法对菌株进行13种抗菌药物的最小抑菌浓度（MIC）检测；采用PCR方法对分离菌株进行耐药基因（bla_KPC-2、bla_NDM-1、bla_NDM-5、bla_CTX-M1、tetA、tetB、sul1、sul2、oqxA、qnrB、aac （6'）-Ib-cr）和毒力基因（rmpA、entB、iucA、iroN、ycfM、fimH、mrkD、wabG）的检测。结果显示，从150份猪肺样本中成功分离鉴定出116株肺炎克雷伯菌，分离率为77.3%；革兰染色镜检菌体呈红色短杆状，荚膜血清分型以K57型（50.9%）为主。药敏试验结果表明，分离株多呈多重耐药表型，对四环素类、β-内酰胺类及磺胺类药物耐药严重，对美罗培南、黏菌素和恩诺沙星敏感。耐药基因检测显示，检出率最高为sul2基因（60.3%）和oqxA基因（59.5%），检出率最低为bla_NDM-₁与bla_NDM-5基因（均为0.9%）。毒力基因检测显示，检出率最高为ycfM基因（87.1%），检出率最低为iroN基因（1.7%）。结果表明，分离自屠宰场的肺炎克雷伯菌检出率较高，多重耐药形势严峻，耐药基因、毒力基因丰富。本研究为猪源肺炎克雷伯菌的防控提供参考依据。

关键词: 肺炎克雷伯菌, 分离鉴定, 耐药性, 耐药基因, 毒力基因

Abstract: To investigate the drug resistance and virulence gene carrying of Klebsiella pneumoniae isolates, sampling investigation and drug resistance analysis were carried out in a slaughterhouse in Guangzhou. Klebsiella pneumoniae was isolated and identified from pig lung samples, and six common capsular serotypes (K1, K2, K5, K20, K54, K57) were identified, and the minimum inhibitory concentration (MIC) of 13 commonly used antibiotics was detected by the agar dilution method. The resistance genes (bla_KPC-2, bla_NDM-₁, bla_NDM-5, bla_CTX-M1, tetA, tetB, sul1, sul2, oqxA, qnrB, aac(6')-Ib-cr) and virulence genes (rmpA, entB, iucA, iroN, ycfM, fimH, mrkD, wabG) were detected by PCR. The results showed that 116 strains of Klebsiella pneumoniae were successfully isolated and identified from 150 pig lung samples, the isolation rate was 77.3%. The bacteria showed red short rod shape by Gram staining microscopy, and serotype of capsule was mainly K57 (50.9%). The results of drug sensitivity test showed that most of the isolates showed multi-drug resistance phenotype, and were highly resistant to tetracycline, β-lactam and sulfonamides, and sensitive to meropenem, colistin and enrofloxacin. The detection of drug resistance genes showed that the highest detection rate was sul2 gene (60.3%) and oqxA gene (59.5%), and the lowest detection rates were bla_NDM-1 and bla_NDM-5 genes (both 0.9%). The virulence gene detection showed that the highest positive rate was ycfM gene (87.1%), and the lowest positive rate was iroN gene (1.7%). The results showed that the detection rate of Klebsiella pneumoniae isolated from slaughterhouse was high, the situation of multiple drug resistance was severe, and the resistance genes and virulence genes were abundant, providing reference for the prevention and control of swine Klebsiella pneumoniae.

Key words: Klebsiella pneumoniae, separation and identification, drug resistance, drug resistance gene, virulence gene

中图分类号:

S852.61

赵菲菲, 李杰, 韩宁, 谢仕廷, 曾振灵. 分离自屠宰场的肺炎克雷伯菌的耐药性分析[J]. 畜牧兽医学报, 2023, 54(7): 3044-3053.

ZHAO Feifei, LI Jie, HAN Ning, XIE Shiting, ZENG Zhenling. Antibacterial Drug Resistance Analysis of Klebsiella pneumoniae Isolated from Slaughterhouse[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 3044-3053.

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