肺炎克雷伯菌超广谱β-内酰胺酶基因型研究  

The genotypes of extended-spectrum beta-lactamases in Klebsiella pneumoniae isolates from a hospital in Kunming

作　　者：胡大春[1] 邵剑春[1] 杨绍敏[1] 周玲[1] 李超[1] 刘德华[1] 

机构地区：[1]昆明市临床疾病分子生物学重点实验室昆明市第一人民医院,昆明650011

出　　处：《中国抗生素杂志》2006年第1期19-22,58,共5页Chinese Journal of Antibiotics

基　　金：昆明市科技局2002年重点科技计划项目(编号:昆科社字2002-01-18)

摘　　要：目的探讨我院肺炎克雷伯菌中超广谱β-内酰胺酶(ESBL s)基因分布规律。方法对20株经双纸片试验确证为ESBL s表型阳性的肺炎克雷伯菌进行blaTEM-1、blaSHV-1、CTX-M-1组、TOHO-1组等4种基因PCR扩增,并对16株blaSHV-1基因PCR扩增阳性的菌株进行基因序列测定,在In ternet网上与G enB ank中的已知序列进行核苷酸相似性分析,并进行编码基因对位和氨基酸序列对比分析。结果产ESBL s肺炎克雷伯菌中blaTEM-1、blaSHV-1、CTX-M-1组等3种基因扩增阳性率分别是50.0%、95.0%、20.0%。16株肺炎克雷伯菌中有4株序列与SHV-1a(序列号:X 98101,74→934)氨基酸序列完全相同;有3株序列与SHV-2(序列号:AY 570959,42→812)100%相同;有2株序列与SHV-11(序列号:AY 293069,41→817)100%相同;有4株序列与SHV-27(序列号:AF 293345,2→821)氨基酸序列完全相同;有1株序列与SHV-28(序列号:AF 538324,12→823)100%相同;有2株序列在G enB ank中未找到与之完全相同的序列。结论本地肺炎克雷伯菌中有SHV-1a、SHV-11、SHV-28广谱β-内酰胺酶基因和SHV-2、SHV-27 ESBL s基因存在。Objective To investigate the genotypes of extended-spectrum＇beta-lactamases （ESBLs） among clinical isolates of Klebsiella pneumoniae. Methods Clinical isolates of Klebsiella pneurnoniae from a hospital in Kunming were screened for ESBL production by disc diffusion methods as described by the CLSI （formerly NCCLS）. ESBL-produeing isolates were confirmed by disc diffusion confirming test as recommended by the CLSI （formerly NCCLS）. PCR analysis was used for blaTEM-1, blaSGW-1, CTX M-1 group and TOHO-1 group genes present in 20 ESBL- producing isolates of Klebsiella pneumoniae. Sequence analysis was performed to identify the blasnv genes in 16 isolates of Klebsiella pneumoniae. Results The genes of blaTEM-1, blaSHV-1, and CTX-M-1 group were detected in 50. 0%, 95.0% and 20.0% of ESBL- producing K. pneumoniae respectively. The ESBL- producing organisms that carried two of these genes was 35.0% of K. pneumoniae. 15.0% of the ESBL- producing K. pneumoniae carried all of the three genes. The TOHO-1 group gene was not detected. Sequence analysis of ESBL- gene showed that there were blaSHV-1, （n= 4）, blaSHV-11 （n = 2）, blaSHV-28 （n = 1）, blaSHV-2（n=3） and blaSHV-27（n=4） in 16 isolates of ESBL- producing K. pneumoniae. There were two sequences of blaSHV-1 gene which was not completely homologous with the genes in GenBank. Conclusion Several blaSHV- genes were detected in clinical isolates of Klebsiella pneumoniae in the hospital in Kunming.

关 键 词：肺炎克雷伯菌 超广谱Β-内酰胺酶 基因 序列分析 

分 类 号：R378[医药卫生—病原生物学]