肺炎克雷伯菌<i>bla</i><sub>CARB-2</sub>基因的分布及结构分析

祝力骋,卢俊婉,王建,许腾,徐娟华

doi:10.16288/j.yczz.18-034

遗传 >

2018 , Vol. 40 >Issue 7: 593 - 600

DOI: https://doi.org/10.16288/j.yczz.18-034

研究报告

肺炎克雷伯菌bla_CARB-2基因的分布及结构分析

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^1. 浙江大学药学院,杭州 310000
^2. 丽水学院医学与健康学院,丽水 323000
^3. 温州医科大学检验医学院,生命科学学院,温州 325000
^4. 宁波市第一医院检验科,宁波 315000

祝力骋,大学,实验师,研究方向：基础医学。E-mail:10594311@qq.com

收稿日期: 2018-03-27

修回日期: 2018-05-31

网络出版日期: 2018-05-31

基金资助

国家自然科学基金(80215049)

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Analyses on distribution and structure of bla_CARB-2 in Klebsiella pneumoniae

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^1. College of Pharmaceutical Sciences,Zhejiang University, Hangzhou 310000, China
^2. College of Medicine and Health, Lishui University, Lishui 323000, China
^3. School of Laboratory and Life Sciences, Wenzhou Medical University, Wenzhou 325000, China
^4. Department of Laboratory Sciences, Ningbo First Hospital, Ningbo 315000, China

Received date: 2018-03-27

Revised date: 2018-05-31

Online published: 2018-05-31

Supported by

Supported by National Natural Science Foundation of China(80215049)

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摘要

为探讨β-内酰胺酶基因在临床分离肺炎克雷伯菌的分布及其相关可移动遗传元件的结构,本文利用基因组测序、PCR、分子克隆、接合转移和基因组学分析等方法,在对240株临床分离的肺炎克雷伯菌进行混合基因组测序的基础上,着重研究了bla_CARB-2等耐药性基因相关可移动遗传元件的结构及其在肺炎克雷伯菌基因组的定位,克隆获得耐药性质粒携带的若干耐药性基因并测定了它们的功能。结果显示,在240株肺炎克雷伯菌中检出11种β-内酰胺酶基因,其中1株肺炎克雷伯菌(KP1276)被检出bla_CARB-2基因,阳性率为0.42% (1/240);bla_CARB-2位于一个大小为182,450 bp的可接合转移质粒(pKP1276-182),pKP1276-182共编码222个基因,包含7个耐药性基因,分别是bla_CARB-2、bla_KLUC、aadA1、aadA2、cmlA1、dfrA1和sul2。bla_CARB-2是首次在肺炎克雷伯菌中被检出,且bla_CARB-2与其他3个耐药性基因一起构成一个新型结构的1型整合子(int-bla_CARB-2-aadA2-cmlA1-aadA1);对其中的3个耐药性基因bla_CARB-2、aadA2和CmlA1进行了分子克隆和耐药性测定,结果显示它们对相应药物都具有一定的耐药性, 其中bla_CARB-2对青霉素类β-内酰胺药物具有较高的耐药性。本研究结果表明,bla_CARB-2基因已经在肺炎克雷伯菌中出现,其由1型整合子携带编码位于一个可接合转移的质粒上,bla_CARB-2基因有可能在相同及不同种属肠杆菌之间进行水平转移,引起耐药性播散。

关键词： 肺炎克雷伯菌; bla_CARB-2; 1类整合子; 克隆; 耐药性

本文引用格式

祝力骋,卢俊婉,王建,许腾,徐娟华 . 肺炎克雷伯菌bla_CARB-2基因的分布及结构分析[J]. 遗传, 2018 , 40(7) : 593 -600 . DOI: 10.16288/j.yczz.18-034

Abstract

In order to characterize the structure of the beta-lactamase gene and its corresponding mobile genetic elements in Klebsiella pneumoniae, the beta-lactamase genes from 240 clinical Klebsiella pneumoniae isolates were studied. bla_CARB-2, a newly characterized gene, was extensively investigated utilizing next-generation sequencing, PCR, molecular cloning, conjugation, and comparative genomics analysis. We identified 11 beta-lactamase genes among the 240 clinical Klebsiella pneumoniae isolates; the bla_CARB-2 gene exists only in one specific isolate (Klebsiella pneumoniae KP1276) (1/240, 0.42%). The bla_CARB-2 gene lies on a conjugative plasmid pKP1276-82, a 182,450-bp plasmid, which encodes 222 open reading frames. The plasmid has seven resistance genes, termed bla_CARB-2, bla_KLUC, aadA1, aadA2, cmlA1, dfrA1, and sul2. Among these genes, bla_CARB-2 was identified for the first time in Klebsiella pneumoniae. Four of these resistance genes and an int gene form a class 1 integron (int-bla_CARB-2-aadA2-cmlA1-aadA1). Further studies show that the bla_CARB-2, aadA2, and cmlA1 genes are resistant to their corresponding antibiotics and the bla_CARB-2 exhibits higher resistance activities to penicillin beta-lactams. These results reveal the possibility of horizontal transfer of the resistance genes and dissemination of resistance among bacteria of different genera or species of Enterobacteriaceae.

Key words： Klebsiella pneumoniae; bla_CARB-2; class 1 integron; cloning; antibiotics resistance

参考文献

[1]

Li B, Zhao Y, Liu C, Chen Z, Zhou D . Molecular pathogenesis of Klebsiella pneumoniae. Future Microbiol, 2014,9(9):1071-1081.

[2]

Swathi CH, Chikala R, Ratnakar KS, Sritharan V . A structural, epidemiological & genetic overview of Klebsiella pneumoniae carbapenemases (KPCs). Indian J Med Res, 2016,144(1):21-31.

[3]

Papadimitriou-Olivgeris M, Marangos M, Christofidou M, Fligou F, Bartzavali C, Panteli ES, Vamvakopoulou S, Filos KS, Anastassiou ED . Risk factors for infection and predictors of mortality among patients with KPC-producing Klebsiella pneumoniae bloodstream infections in the intensive care unit. Scand J Infect Dis, 2014,46(9):642-648.

[4]

Karaiskos I, Giamarellou H . Multidrug-resistant and extensively drug-resistant Gram-negative pathogens: current and emerging therapeutic approaches. Expert Opin Pharmacother, 2014,15(10):1351-1370.

[5]

Tangdén T, Giske CG . Global dissemination of extensively drug-resistant carbapenemase-producing Enterobacteriaceae: clinical perspectives on detection, treatment and infection control. J Intern Med, 2015,277(5):501-512.

[6]

Robilotti E, Deresinski S . Carbapenemase-producing Klebsiella pneumoniae. F1000Prime Rep, 2014,6:80.

[7]

Clinical and Laboratory Standards Institute (CLSI). Performance standards for antimicrobial susceptibility testing; 27th informational supplement.Standard: CLSI-M100- S27.

[8]

J.萨姆布鲁克(SambrookJ.), D.W.拉塞尔. 分子克隆实验指南(第三版). 黄培堂等译, 2002, 北京: 科学出版社.

[9]

Delcher AL, Bratke KA, Powers EC, Salzberg SL . Identifying bacterial genes and endosymbiont DNA with Glimmer. Bioinformatics, 2007,23(6):673-679.

[10]

Posada D. Using MODELTEST and PAUP* to select a model of nucleotide substitution. Curr Protoc Bioinformatics, 2003, Chapter 6: Unit6.5.

[11]

Xu T, Ying J, Yao XD, Song YL, Ma P, Bao BK, Jiang WY, Wu XM, Tou HF, Li PZ, Ren P, Fei JX, Yang L, Liu Q, Xu ZY, Zhou TL, Ni LY, Bao QY . Identification and characterization of two novel bla(KLUC) resistance genes through large-scale resistance plasmids sequencing. PLoS One, 2012,7(10):e47197.

[12]

Rice LB, Carias LL, Bonomo RA, Shlaes DM . Molecular genetics of resistance to both ceftazidime and beta-lactam- beta-lactamase inhibitor combinations in Klebsiella pneumoniae and in vivo response to beta-lactam therapy. J Infect Dis, 1996,173(1):151-158.

[13]

Zhang YJ, Chang QL, Wang Q, Lu JW, Wang H, Li PZ, Ying J, Bao QY, Hu YL . Structure and function of class 1 integron in clinical isolates of Klebsiella pneumoniae. Hereditas (Beijing), 2014,36(6):603-610.

[13]

张跃进, 常清利, 汪倩, 卢俊婉, 王欢, 李佩珍

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