多药耐药大肠杆菌OXA基因检测及聚类分析  

Detection of Multi-drug Resistant Escherichia Coli OXA Gene and Cluster Analysis

作　　者：胡海莹[1] 王冬国[2] 魏志英[1] 郑雨薇[1] 陈玲敏[1] 方小芳[1] 戚永孝[1] 

机构地区：[1]浙江省台州学院医学院检验系,318000 [2]浙江省台州学院医学院附属市立医院检验科,318000

出　　处：《医学研究杂志》2012年第9期129-133,共5页Journal of Medical Research

基　　金：浙江省大学生科技创新活动计划(新苗人才计划)(2010R428027)

摘　　要：目的研究笔者医院多药耐药的大肠杆菌OXA基因的分布特性。方法采用MIC法,用Microscan公司肉汤稀释法药敏板来检测细菌的药敏结果;PCR检测OXA-1 Cluster、OXA-2 Cluster、OXA-10 Cluster、OXA-20 Cluster、OXA-23 Clus-ter、OXA-24 Cluster、OXA-51 Cluster、OXA-58 Cluster等基因,阳性产物进行DNA测序以确定基因亚型;用SPSS 17.0软件作聚类分析。结果药敏试验结果,实验菌株对阿米卡星、头孢西丁、头孢哌酮/舒巴坦、哌拉西林/他唑巴坦的敏感率为60%～70%,对亚胺培南、美洛培能的敏感率为100%,而对头孢菌素类抗生素的敏感率较低,在30%以下;OXA基因PCR结果:以OXA-10 cluster和OXA-2 cluster流行为主,分别占检测菌株的31.3%和28.1%;测序结果:在OXA-10 cluster中以OXA-10基因与OXA-17为主,分别占3/10(30.0%),OXA-16基因次之,占2/10(20.0%);在OXA-2 cluster中以OXA-15基因为主,占4/9(44.4%),OXA-3基因和OXA-53基因次之,分别占2/9(22.2%);对分别以1,2,3,…,31,32编号的32株实验菌株OXA基因的检测结果聚类分析结果表明:实验菌株28-32-6-23-26-18-22-12-15-11株,8-30株,17-31-3株,1-24株,19-27-2-10-14-5株,4-29株,7-1株为OXA耐药基因克隆传播菌株;进一步对OXA基因测序结果作聚类分析结果表明:OXA-25、OXA-33、OXA-4、OXA-13、OXA-56基因型,OXA-10、OXA-17基因型,OXA-16、OXA-24、OXA-1、OXA-3、OXA-53基因型,OXA-15、OXA-20基因型为OXA耐药基因克隆传播的类型。结论在笔者医院存在OXA耐药基因医院内感染的趋势,并有暴发性流行的可能。Objective To study characteristics of distribution of multi - drug resistant Escherich^a coli OXA gene in our hospital. Methods Using MIC method to detect susceptibility with Microscan＇s broth dilution plate;using PCR to deteete OXA - 1 Cluster, OXA -2 Cluster, OXA- 10 Cluster, OXA-20 Cluster, OXA-23 Cluster, OXA-24 Cluster, OXA-51 Cluster, OXA-58 Cluster genes. DNA sequencing was used to determine the genetic subtypes of positive products; Using SPSS 17.0 software to do cluster analysis. Re- sults Sensitivity rates about Amikacin, Cefoxitin, Cefoperazone/Sulbactam and Piperacillin/Tazobactam were 60% -70% , sensitivity rates about Imipenem and Meropnem were 100%. However, sensitivity rates about Cephalosporins are lower（ 〈 30% ） ; the results of OXA gene PC R：OXA - 10 cluster and OXA -2 cluster wethe main prevalent genes and they accounted for 31.3 % and 28.1% , separate- ly; the results of sequencing： OXA- 10 and OXA -17 were the mainegenes in OXA- 10 cluster,and they accounted for 3/10（ 30.0% ） , separately, following was OXA - 16, which accounted for 2/10（20.0% ） ; It is main gene for OXA - 15 was the main gene in OXA -2 cluster, which accounted for 3/10 （30.0%） , following were OXA - 3 and OXA - 53, and they accounted for 2/9 （22.2%） ; the result of cluster analysis： the tested isolates about 28 -32 -6 -23 -26 - 18 -22 - 12 - 15 - 11,8 -30,17 -31 -3,1 -24,19 - 27 -2 - 10 - 14 -5,4 -29,7 - 13 wer cloning spreading for OXA resistant isolates;Further sequencing of the OXA genes for clustering analysis showed： OXA-25, OXA-33, OXA-4, OXA-13, OXA-56 genotype, OXA-10, OXA-17 genotype, OXA-16, OXA -24, OXA-1, OXA- 3, OXA- 53 genotype, OXA -15, OXA -20 were cloning spreading for OXA resistant genotypes. Conclusion There exists trend of nosoeomial infection and outbreaks of epidemic in our hospital.

关 键 词：多药耐药 大肠杆菌 OXA基因 聚类分析 

分 类 号：R512.62[医药卫生—内科学]