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烧伤患者碳青霉烯类耐药菌耐药机制及毒力特征的初步研究

[Preliminary study on resistance mechanism and virulence features in carbapenems-resistant from burn patients].

作者信息

Chen Z, Xiang J

机构信息

Department of Burns and Plastic Surgery, Ruijin Hospitals, School of Medicine, Shanghai Jiao Tong University, Shanghai 200025, China.

出版信息

Zhonghua Shao Shang Za Zhi. 2018 Nov 20;34(11):796-801. doi: 10.3760/cma.j.issn.1009-2587.2018.11.015.

Abstract

To investigate resistance mechanism and virulence features of carbapenems-resistant (CRKP) and to provide theoretical foundation for the prevention and control of nosocomial infections in burn wards as well as clinical therapy of CRKP. Strains of isolated from clinical samples of burn patients hospitalized in our unit from February to December 2016 were collected. The diameters of inhibition zone of imipemen or meropenem susceptibility disk of the above strains were≤22 mm. Kirby-Bauer disk diffusion method and automated instrument were applied to test drug resistance of the strains to 17 antibiotics of cephalosporins, aminoglycosides, sulfonamides, quinolones, and carbapenems. Pulsed-field gel electrophoresis (PFGE) was used to analyze homology of the strains. Crystal violet staining was applied to assess ability of biofilm formation of the strains. The hypermucoviscosity (HM) phenotype of the strains was determined by string test. Polymerase chain reaction was used to detect common carbapenemase genes of (KPC), (IMP), (OXA-48), and (NDM), capsular serotype genes of 1, 2, and 57, and virulence-associated gene of rmpA. (1) A total of 29 CRKP strains were isolated. The 29 CRKP strains were with high drug resistance rate to carbapenems antibiotics of imipenen, meropenem, and ertapenem, and cephalosporins, aminoglycosides, and quinolones antibiotics, while the strains were with low drug resistance rate to sulfonamides antibiotics and tegafycline. (2) The 29 CRKP strains had 4 types of A, B, C, and D according to PFGE, with 11 strains of type A, 10 strains of type B, 6 strains of type C, and 2 strains of type D. (3) Among the 29 CRKP strains, 25 strains were positive in biofilm formation, with positive rate of 86.2%. (4) None of the 29 CRKP strains was positive in the string test with HM phenotype. (5) All the 29 CRKP strains carried carbapenemase genes. Among the 29 CRKP strains, 11 strains carried both (NDM-1) and (OXA-48) genes, 1 strain carried both (NDM-1) and (KPC-2) genes, 12 strains only carried (KPC-2) gene, and 5 strains only carried (NDM-1) gene. The 79.3% (23/29) strains were completely resistant to 3 kinds of carbapenems antibiotics. (6) Among the 29 CRKP strains, 3 strains carried 2 gene, 2 of which were completely resistant to 3 kinds of carbapenems antibiotics, 1 strain carried gene, and no strain carried 1 or 57 gene. The detection rate of CRKP isolated from burn patients hospitalized in our unit is high. The strains are mostly extensively drug-resistant bacteria, and the mechanism of drug resistance of which is complicated. There may have clonal spread of CRKP. And appropriate measures should be taken timely and effectively to prevent nosocomial spread of CRKP.

摘要

探讨耐碳青霉烯类肺炎克雷伯菌(CRKP)的耐药机制及毒力特征,为烧伤病房医院感染防控及CRKP的临床治疗提供理论依据。收集2016年2月至12月在我科住院的烧伤患者临床标本分离的菌株。上述菌株亚胺培南或美罗培南药敏纸片抑菌圈直径≤22 mm。采用 Kirby-Bauer 纸片扩散法及自动化仪器检测菌株对头孢菌素类、氨基糖苷类、磺胺类、喹诺酮类及碳青霉烯类17种抗生素的耐药情况。采用脉冲场凝胶电泳(PFGE)分析菌株同源性。采用结晶紫染色法评估菌株生物膜形成能力。采用悬滴试验检测菌株的高黏液性(HM)表型。采用聚合酶链反应检测常见碳青霉烯酶基因(KPC)、(IMP)、(OXA-48)和(NDM)、荚膜血清型基因1、2和57以及毒力相关基因rmpA。(1)共分离出29株CRKP菌株。29株CRKP菌株对亚胺培南、美罗培南和厄他培南等碳青霉烯类抗生素以及头孢菌素类、氨基糖苷类和喹诺酮类抗生素耐药率高,而对磺胺类抗生素和替加环素耐药率低。(2)29株CRKP菌株根据PFGE分为A、B、C、D 4型,其中A型11株,B型10株,C型6株,D型2株。(3)29株CRKP菌株中,25株生物膜形成阳性,阳性率为86.2%。(4)29株CRKP菌株悬滴试验HM表型均为阴性。(5)29株CRKP菌株均携带碳青霉烯酶基因。29株CRKP菌株中,11株同时携带(NDM-1)和(OXA-48)基因,1株同时携带(NDM-1)和(KPC-2)基因,12株仅携带(KPC-2)基因,5株仅携带(NDM-1)基因。79.3%(23/29)的菌株对3种碳青霉烯类抗生素完全耐药。(6)29株CRKP菌株中,3株携带2型基因,其中2株对3种碳青霉烯类抗生素完全耐药,1株携带型基因,未发现携带1或57型基因的菌株。从我科住院烧伤患者中分离出CRKP的检出率较高。菌株多为广泛耐药菌,耐药机制复杂。CRKP可能存在克隆传播。应及时有效采取相应措施预防CRKP的医院内传播。

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