283株肠杆菌科产ESBLs菌的基因型分析  

Genotypes of Extended-Spectrum β-Lactamases in Clinical Isolates of Enterobacteriaceae

作　　者：张亚兰 王骋 胡佳杰[3] 

机构地区：[1]武汉市第五医院检验科,武汉430050 [2]马应龙药业集团股份有限公司,武汉430064 [3]湖北中医药大学检验学院,武汉430065

出　　处：《现代检验医学杂志》2013年第5期60-64,共5页Journal of Modern Laboratory Medicine

摘　　要：目的 了解武汉市第三医院临床分离的肠杆菌科细菌中产超广谱β-内酰胺酶菌株的基因型及感染现状.方法 收集武汉市第三医院2012年1月～12月临床分离的双纸片协同初筛试验为阳性的产ESBLs肠杆菌科细菌,采用临床实验室标准化协会推荐的标准纸片扩散法表型确证试验确证ESBLs阳性菌,采用PCR技术以及基因测序技术对确证试验为阳性的产ESBLs菌进行基因分型.结果 共收集333株ESBLs初筛试验阳性的肠杆菌科细菌,运用标准纸片扩散法表型确证试验确证ESBLs阳性菌283株,其中大肠埃希菌190株,肺炎克雷伯菌66株,奇异变形杆菌10株,普通变形杆菌6株,阴沟肠杆菌11株.283株产ESBLs菌中,66株TEM-1基因阳性（23.32%）,0株SHV基因阳性（0%）,103株CTX-M-15基因阳性（36.40%）,3株CTX-M-3基因阳性（1.06%）,2株CTX-M-14基因阳性（0.71%）,45株菌TEM-1和CTX-M-15两种基因型同时阳性（15.90%）,64株细菌未检出ESBLs基因（22.61%）.结论 该院最为流行的产ESBLs病原菌是大肠埃希菌和肺炎克雷伯菌,产ESBLs肠杆菌科细菌中携带多种β-内酰胺酶基因,较为流行的耐药基因型是CTX-M-15和TEM-1.Objective To investigate the main genotypes and the epidemic characteristics of ESBLs isolates in the Third Hos-pital of Wuhan City. Methods The ESBLs phenotype of 333 clinical isolates of Enterobacteriaceae was primarily screened and the isolates were collected in the third hospital of Wuhan city from January to December 2012. The ESBLs phenotype of clinical isolates were confirmed by CLSI phenotypic confirmatory test and according to guidelines of CLSI. ESBLs phenotype-positive clinical isolates using primers specific for TEM, CTX-M and SHV respectively, and the polymerase chain reaction （PCR） products were purified and sequenced. The sequenced fragments will be blasted with NCBI gene bank for ESBLs gen- otypes. Results 283 strains were ESBLs producing positive strains by confirmatory test,including 190 Escherichia coli ,66 Klebsierllapneurnoniae,10 Proteus rnirabilis, 6 Proteus vulgaris and 11 Enterobacter cloacae. CTX-M-15 accounted for 36.40% （103/283） ,CTX-M-14 accounted for 0.71% （2/283） ,CTX-M-3 accounted for 1.06% （3/283） ,TEM-1 type accoun- ted for 23.32 % （66/283）, 45 （15.90%） were both blaTEM genes and balCTX-M genes, 22.61% （64/283） of the ESBLs phenotype positive clinical isolates had not been amplificated to be positive strip. SHV genotype was not found. Conclusion The experimental results showed the most common ESBLs producing isolates were Escherichia coli and Klebsierlla pneu-moniae, ESBLs producing enterobacteriaceae bacteria carried in a variety of beta-lactamase gene. The highest prevalence re-sistance genetype was bla CTX-M-15 gene.

关 键 词：超广谱Β-内酰胺酶 耐药基因 聚合酶链反应 基因分型 

分 类 号：R378.2[医药卫生—病原生物学] R446.7[医药卫生—基础医学]