Department of Clinical Laboratory Science, Dong-eui University, Busan, 614-714, Republic of Korea.
J Microbiol. 2010 Jun;48(3):358-64. doi: 10.1007/s12275-010-9308-5. Epub 2010 Jun 23.
This study was performed to characterize the chromosomal metallo-beta-lactamases (MBLs) of Elizabethkingia meningoseptica isolated from Korea and to propose a clustering method of BlaB and GOB MBLs based on their amino acid similarities. Chromosomal MBL genes were amplified by PCR from 31 clinical isolates of E. meningoseptica. These PCR products were then cloned into a vector and electrotransformed into E. coli DH5 alpha. Nucleotide sequencing was performed by the dideoxy chain termination method using PCR products or cloned DNA fragments. Antimicrobial susceptibilities were determined by the agar dilution method. PCR experiments showed that all 31 E. meningoseptica isolates contained both the blaB and the bla (GOB) genes. DNA sequence analysis revealed that E. meningoseptica isolates possessed seven types of blaB gene, including five novel variants (blaB-9 to blaB-13) and 11 types of bla (GOB) gene, including 10 novel variants (bla (GOB-8) to bla (GOB-17)). The most common combination of MBL was BlaB-12 plus GOB-17 (n=19). Minimum inhibitory concentrations of imipenem and meropenem for the electrotransformants harboring novel BlaB and GOB MBLs were two- or four-fold higher than those for the recipient E. coli DH5 alpha. BlaB and GOB MBLs were grouped in three and six clusters including fifteen novel variants, respectively, based on amino acid similarities.
本研究旨在对分离自韩国的脑膜败血伊丽莎白菌的染色体金属β-内酰胺酶(MBLs)进行特征分析,并提出一种基于氨基酸相似性的 BlaB 和 GOB MBLs 聚类方法。采用 PCR 从 31 株临床分离的脑膜败血伊丽莎白菌中扩增染色体 MBL 基因。将这些 PCR 产物克隆到载体中,然后电转化到大肠杆菌 DH5α中。采用 PCR 产物或克隆 DNA 片段的双脱氧链终止法进行核苷酸测序。采用琼脂稀释法测定抗菌药物敏感性。PCR 实验表明,31 株脑膜败血伊丽莎白菌均含有 blaB 和 bla(GOB)基因。DNA 序列分析显示,脑膜败血伊丽莎白菌分离株携带 7 种 blaB 基因,包括 5 种新的变异体(blaB-9 至 blaB-13)和 11 种 bla(GOB)基因,包括 10 种新的变异体(bla(GOB-8)至 bla(GOB-17))。最常见的 MBL 组合是 BlaB-12 加 GOB-17(n=19)。携带新型 BlaB 和 GOB MBL 的电转化子对亚胺培南和美罗培南的最小抑菌浓度比受体大肠杆菌 DH5α高出两到四倍。基于氨基酸相似性,BlaB 和 GOB MBL 分别分为三组和六组,包括 15 种新型变异体。