Hu Gong-Zheng, Chen Hong-Ying, Si Hong-Bin, Deng Li-Xin, Wei Zhan-Yong, Yuan Li, Kuang Xiu-Hua
Department of Pharmacology and Toxicology, College of Animal Husbandry and Veterinary Science, Henan Agricultural University, Zhengzhou, China.
FEMS Microbiol Lett. 2008 Feb;279(2):162-6. doi: 10.1111/j.1574-6968.2007.01017.x. Epub 2007 Dec 18.
Extended-spectrum beta-lactamases (ESBLs) produced by a clinical isolate of Shigella flexneri from chickens were detected with confirmatory phenotypic tests of the Clinical and Laboratory Standards Institute, and minimum inhibitory concentrations of several antibacterial drugs against the isolate were determined by the twofold dilution method. The genotype and subtype of the ESBL-producing S. flexneri isolate were identified by PCR amplifying of ESBL genes and DNA sequencing analysis. The results revealed that the isolate was able to produce ESBLs. They were resistant to third-generation cephalosporins such as ceftiofur and ceftriaxone and showed characteristics of multidrug resistance. The ESBL gene from the S. flexneri isolate was of the TEM type. Sequence analysis indicated that the TEM-type gene had 99.1% and 99.2% identity to TEM-1D ESBL and TEM-1 beta-lactamase, respectively, at the nucleotide level. The amino acid sequence inferred from the TEM-type gene revealed three substitutions compared with the TEM-1 and TEM-1D enzymes: Ser51Gly, Val82Ila and Ala182Val. When it was compared with TEM-116 (99.8% identity), there were only two mutations (A(151)G and T(403)C) in the TEM-type gene, resulting in the substitution of Ser to Gly at position 51 in the amino acid sequence. The TEM type was a TEM-116 derivative.
通过临床和实验室标准协会的确认性表型试验,检测了从鸡中分离出的弗氏志贺氏菌临床分离株产生的超广谱β-内酰胺酶(ESBLs),并通过两倍稀释法测定了几种抗菌药物对该分离株的最低抑菌浓度。通过对ESBL基因进行PCR扩增和DNA测序分析,鉴定了产生ESBL的弗氏志贺氏菌分离株的基因型和亚型。结果显示,该分离株能够产生ESBLs。它们对头孢噻呋和头孢曲松等第三代头孢菌素耐药,并表现出多重耐药特征。弗氏志贺氏菌分离株的ESBL基因属于TEM型。序列分析表明,该TEM型基因在核苷酸水平上与TEM-1D ESBL和TEM-1β-内酰胺酶的同一性分别为99.1%和99.2%。从TEM型基因推断的氨基酸序列与TEM-1和TEM-1D酶相比,有三个替换:Ser51Gly、Val82Ila和Ala182Val。当与TEM-116(同一性为99.8%)比较时,TEM型基因中只有两个突变(A(151)G和T(403)C),导致氨基酸序列中第51位的Ser被Gly取代。该TEM型是TEM-116的衍生物。