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血流感染碳青霉烯耐药肺炎克雷伯菌的耐药基因和毒力基因及分子流行病学研究

Carbapenemase Genes, Virulence Genes, and Molecular Epidemiology of Carbapenem-Resistant Klebsiella pneumoniae Derived From Bloodstream Infections

  • 摘要:
    目的 本研究旨在了解西南地区某大型三甲综合医院血流感染碳青霉烯耐药肺炎克雷伯菌(carbapenem-resistant Klebsiella pneumoniae, CRKP)患者的临床特征和CRKP的分子流行病学。
    方法 收集2015–2019年血流感染患者血培养分离的131株非重复CRKP,采用VITEK-2全自动微生物分析仪和质谱MALDI-TOF进行菌株鉴定,微量肉汤稀释法检测最低抑菌浓度值(minimum inhibitory concentration, MIC),PCR检测常见碳青霉烯酶耐药基因和毒力因子,多位点序列分型对菌株进行同源性分析,全基因组测序方法检测不携带碳青霉烯酶菌株的基因组特征。
    结果 131株CRKP除多粘菌素B(耐药率1.6%)和替加环素(耐药率8.0%)外,对常见抗菌药物均耐药。105株(80.2%)CRKP携带Klebsiella pneumoniae carbapenemase (KPC)耐药基因,15株(11.4%)携带New Delhi Metallo-β-lactamase (NDM)基因,4株(3.1%)同时携带KPCNDM基因,以序列(sequence typing, ST)11(74.0%)为优势序列类型,mrkD(96.2%)、fimH(98.5%)、entB(100%)等毒力基因检出率较高,检出1株高毒力CRKP。全基因组测序显示7株不产碳青霉烯酶的CRKP携带ESBLAmpC基因,同时存在膜孔蛋白OMPK35和OMPK36的异常。
    结论 某大型三甲综合医院CRKP主要携带KPC基因,对多种抗菌药物耐药率高,同时携带多种毒力基因,具有高毒力特性的CRKP应引起重视。

     

    Abstract:
    Objective To investigate the clinical characteristics and molecular epidemiology of carbapenem-resistant Klebsiella pneumoniae (CRKP) isolated from patients with bloodstream infections in a large tertiary-care general hospital in Southwest China.
    Methods A total of 131 strains of non-repeating CRKP were collected from the blood cultures of patients who had bloodstream infections in 2015-2019. The strains were identified by VITEK-2, a fully automated microbial analyzer, and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. The minimum inhibitory concentration (MIC) was determined by microbroth dilution method. The common carbapenemase resistant genes and virulence factors were identified by PCR. Homology analysis was performed by multilocus sequencing typing. Whole genome sequencing was performed to analyze the genomic characteristics of CRKP without carbapenemase.
    Results The 131 strains of CRKP showed resistance to common antibiotics, except for polymyxin B (1.6% resistance rate) and tigacycline (8.0% resistance rate). A total of 105 (80.2%) CRKP strains carried the Klebsiella pneumoniae carbapenemase (KPC) resistance gene, 15 (11.4%) strains carried the New Delhi Metallo-β-lactamase (NDM) gene, and 4 (3.1%) isolates carried both KPC and NDM genes. Sequence typing (ST) 11 (74.0%) was the dominant sequence type. High detection rates for mrkD (96.2%), fimH (98.5%), entB (100%), and other virulence genes were reported. One hypervirulent CRKP strain was detected. The seven strains of CRKP that did not produce carbapenemase were shown to carry ESBL or AmpC genes and had anomalies in membrane porins OMPK35 and OMPK36, according to whole genome sequencing.
    Conclusion In a large-scale tertiary-care general hospital, CRKP mainly carries the KPC gene, has a high drug resistance rate to a variety of antibiotics, and possesses multiple virulence genes. Attention should be paid to CRKP strains with high virulence.

     

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