β-内酰胺酶TEM、SHV全长基因检测与应用

Detection and Use of TEM and SHV Full Length Genes of β-Lactamase

目的 建立 β-内酰胺酶 TEM、SHV全长基因检测方法 ,并对临床分离株进行检测分析。方法 检索 Gen-Bank核酸库中已登录的 TEM、SHV各亚型基因序列 ,在保守区分别设计引物、分段扩增、分段测序 ,并由测序仪软件拼接成全长基因序列。结果 大肠埃希菌 TEM- 1型、TEM- 2型、TEM- 6型、TEM- 2 6型、SHV- 1型、SHV- 2型测得序列与期望序列一致 ;从大肠埃希菌 (苏州 )、肺炎克雷伯菌 (苏州 )、铜绿假单胞菌 (温州 )、鲍氏不动杆菌(湖州 )、鲁氏洛菲不动杆菌 (济南 )、褪色 (粘质 )沙雷菌 (济南 )、流感嗜血菌 (苏州 )、副流感嗜血菌 (苏州 )、肺炎链球菌 (苏州 )、淋病奈瑟球菌 (常州 )、MRSA(常州 )、MRCNS(鄂尔多斯 )等菌中检出 TEM序列 ,并有 4条 TEM序列登录 Gen Bank;从铜绿假单胞菌 (温州 )、鲍氏不动杆菌 (湖州 )等菌中检出 SHV序列 ,并有 3条 SHV序列登录Gen Bank。结论 本 β-内酰胺酶的 TEM。

OBJECTIVE To set up the detection methods of TEM and SHV full length genes of β-lactamase, to detect and analyze the clinical isolated strains. METHODS Every second model genes sequences of TEM and SHV of registered GenBank were searched. In the conserved region, design of primers and amplification of divided section and detection sequences of divided section were individually carried out. It was performed by software of instrument of detection sequences that put full length genes together. RESULTS The detected sequences were the same as predicted in TEM-1 type, TEM-2 type, TEM-6 type, TEM-26 type, SHV-1 type and SHV-2 type of Escherichia coli; the sequences of TEM were positive from the detection of E.coli (Suzhou), Klebsiella pneumoniae (Suzhou), Pseudomonas aeruginosa (Wenzhou), Acinetobacter baumannii (Huzhou), A. lwoffi (Jinan), Serratia marcescens (Jinan), Haemophilus influenzae (Suzhou), H. parainfluenzae (Suzhou), Streptococcus pneumoniae (Suzhou), Neisseria gonorrhoeae (Changzhou), MRSA (Changzhou) and MRCNS (Erduosi), 4 strip sequences of TEM were registered on GenBank; the sequences of SHV were positive from the detection of P. aeruginosa (Wenzhou), A. baumannii (Huzhou), 3 strip sequences of SHV were registered on GenBank. CONCLUSIONS The detection methods of TEM and SHV full length genes of β-lactamase could obtain a good result in practical use.