高耐药洛菲不动杆菌分离株超广谱β内酰胺酶的基因变异

山东大学学报(医学版)

高耐药洛菲不动杆菌分离株超广谱β内酰胺酶的基因变异

于秀娟¹,姚苹²,孔庆莲¹

山东大学 1.齐鲁医院检验科； 2.公共卫生学院卫生检验研究室，山东济南 250012

收稿日期:2005-12-08 修回日期:1900-01-01 出版日期:2007-04-24 发布日期:2007-04-24
通讯作者: 于秀娟

Differentiation of bla-TEM genes in highly resistant Acinetobacter lwoffi strains

YU Xiu-juan¹,YAO Ping²,KONG Qing-lian¹

1. Clinical Laboratory, Qilu Hospital of Shandong University; 2. Department of Public Health Laboratory,School of Public Health, Shandong University

Received:2005-12-08 Revised:1900-01-01 Online:2007-04-24 Published:2007-04-24
Contact: YU Xiu-juan

摘要/Abstract

摘要： 目的：探讨高度耐药的洛菲不动杆菌超广谱β内酰胺酶基因型及基因变异特征，分析基因变异对氨基酸序列及对超广谱β内酰胺酶可能产生的影响。方法：从下呼吸道感染的患者标本中分离得到3株高度耐药的洛菲不动杆菌超广谱β内酰胺酶阳性菌，PCR检测TEM、SHV、OXA、IMP、CTX-M耐药基因，并对耐药基因进行全序列测定，比较不同菌株耐药基因及其编码氨基酸序列的变异特征。结果：2株耐药基因为TEM型，SHV、OXA、IMP、CTX-M耐药基因均未检出，另1株5种耐药基因均未检出。2株TEM基因扩增序列分别长1012bp、1007bp，2株同源性为98.2%，在17个位点上存在碱基差异，TEM基因为832bp，编码277个氨基酸，两株间有8个氨基酸出现差异，差异主要位于天冬氨酸、甘氨酸和丝氨酸，这些氨基酸分别被组氨酸、精氨酸/丙氨酸、半胱氨酸取代。软件分析显示，2株TEM基因编码TEM型超广谱β内酰胺酶的等电点分别为5.479、5.083。结论：山东济南地区高度耐药洛菲不动杆菌ESBLs耐药基因可见TEM型，不同临床分离株耐药基因存在不同程度的变异，这种变异使其编码的氨基酸序列发生改变，可能影响超广谱β内酰胺酶的某些特性，给耐药菌的检测带来影响。

Abstract: Objective: To discover the extended spectrum beta-lactamases (ESBLs) gene types and the characteristics of Acinetinbacter lwoffi strains which were highly resistant to most kinds of antibiotics and to compare the differentiation in nucleotide and amino acid sequences to show the possible influence in laboratory. Methods: Three strains of A.lwoffi with ESBLs phenomenon were isolated from patients′ sputum suffering from respiratory infection. The genes of blaTEM, SHV, OXA, IMP and CTX-M were determined by PCR. Nucleotide sequences were detected by ABI automated sequencer and analyzed by the DNA star software to compare the differentiation in nucleotide and amino acid sequences. Results: The bla-TEM gene type was determined in two strains, but the blaTEM, SHV, OXA, IMP and CTX-M genes were not determined in another strain. Amplified nucleotide sequences of TEM genes were 1012bp and 1007bp, respectively. The identical rate was 98.2%. There were 17 pairs different in them. The blaTEM gene, 832bp, coded for 277 amino acids. Among them, 8 amino acids were different, which mainly happened on Asp, Gly and Ser, substituted by His, Arg/Ala and Cys. They had pI value of 5.479 and 5.083, respectively. Conclusion: The blaTEM gene presented in A.lwoffi highly resistant strains in Shandong province. However, they produce some differentiations in nucleotide and amino acid sequences. These differentiations influence some conditions of laboratory method to pick up the resistant strains.

Key words: Acinetobacter lwoffi, extended spectrum beta-lactamases, nucleotide, amino acid, differentiation

中图分类号:

R378

于秀娟,姚苹,孔庆莲 . 高耐药洛菲不动杆菌分离株超广谱β内酰胺酶的基因变异[J]. 山东大学学报(医学版), 2007, 45(4): 384-388.

YU Xiu-juan,YAO Ping,KONG Qing-lian. Differentiation of bla-TEM genes in highly resistant Acinetobacter lwoffi strains[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2007, 45(4): 384-388.

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