鸡大肠杆菌TEM和CTX-M型超广谱β-内酰胺酶基因分型研究.pdfVIP

中国农业科学 2010,43(20):4310-4316 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2010.20.024 鸡大肠杆菌TEM和CTX-M型超广谱 β-内酰胺酶基因分型研究 苑 丽，刘建华，胡功政，潘玉善，莫 娟，魏永俊，裴亚玲 （河南农业大学牧医工程学院，郑州 450002 ） 摘要: 【目的】检测鸡大肠杆菌超广谱β-内酰胺酶（ESBLs）的基因型和基因亚型，了解河南省产酶鸡大肠 杆菌 ESBLs 基因型分布。【方法】 对河南省 7 个地区不同鸡场临床分离 28 株产 ESBLs 鸡大肠杆菌分别用 TEM、 SHV和CTX-M 等3种通用引物进行PCR扩增及测序分析，确定其基因型和基因亚型。【结果】 21 株鸡大肠杆菌检 出TEM型基因，2株检出CTX-M 型基因，5株检出TEM型和CTX-M型两种基因，未检出SHV型。TEM基因亚型以TEM-1 变异型为主，与AY293072（TEM-1）相比同源性为98%—99%，核苷酸序列除发生18T→C 沉默突变外，尚有3株分 别有另一处碱基发生变化：783G→A（C3菌，序列号为FJ405207）、21T→A（X2菌，序列号为FJ405208）和269G →A（F4 菌，序列号为 FJ405211），其中前两处为沉默突变，F4 菌的碱基突变引起 92 甘氨酸变为天冬氨酸，为 TEM-57型。CTX-M基因亚型包括两种：CTX-M-65亚型4株（C2、C3、C4和K1，序列号分别为FJ405191，FJ405192， FJ405212 和 FJ405214）和 CTX-M-14 亚型 3 株（F2、X3 和 B5，其中 F2 菌的相关序列已上传至 GenBank，获序列 号为FJ405213）。【结论】 目前河南省产ESBL鸡大肠杆菌基因亚型以TEM-1变异型为主，CTX-M-65和 CTX-M-14 型次之。 关键词：鸡大肠杆菌；超广谱β-内酰胺酶；TEM基因；CTX-M基因 Genotypes of TEM-Type and CTX-M-Type Extended-Spectrum β-Lactamases Produced by Escherichia coli Isolated from the Fowl YUAN Li, LIU Jian-hua, HU Gong-zheng, PAN Yu-shan, MO Juan, WEI Yong-jun, PEI Ya-ling (College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002) Abstract: 【Objective 】 The genotypes and subtypes of ESBLs produced by Escherichia coli isolated from the fowl were detected to elucidate their distributions in Henan Province. 【Method 】 Twenty-eight ESBL-producing Escherichia coli were collected from different chicken farms of seven districts in Henan Province, and were amplified by PCR using the primers of TEM, SHV and CTX-M, respectively. Their PCR products were cloned and then the cloned fragments were sequenced. The nucleotide sequences were analyzed by DNAStar software and GenBank