超薄层分析性等电聚焦电泳技术在β-内酰胺酶检测中的应用  

DENTIFICATSON OF β-LACTAMASES BY ULTRATHIN-LAYER ISOELECTRIC FOCUSING ELECTROPHORESIS

作　　者：凌宝东 王浴生[2] 刘定勇 

机构地区：[1]华西医科大学基础医学院药理室 [2]华西医科大学临床药理研究所

出　　处：《中国抗生素杂志》1990年第3期184-187,共4页Chinese Journal of Antibiotics

摘　　要：采用“拍板技术”(Flap technique),自制超薄层(0.2mm)聚丙烯酰胺凝胶板.检测了九株Ceftazidime耐药细菌的β-内酰胺酶,并与标准染色体介导的β-内酰胺酶进行比较。除R27853具有两条酶带外,其余均为一条酶条。R1006,R8701,R884三种酶的等电点(pI)与标准酶D-31一致,pI为8.6;R222,R1029两种酶的pI与标准酶P99吻合,pI为8.0。同时,还发现了pI为8.2的R27853和pI为8.4的R471、R855、R875四种酶,目前尚未见文献报道。这一超薄层凝胶技术具有操作简便、快速、灵敏、省时,便于同步比较多个样品及实验结果更易保存等优点,可用作β-内酰胺酶的鉴定与分类,以及进行蛋白质成份分析的方法。Ultrathin polyacrylamide gels(0.2mm) were prepared by the new 'flap technique'.Through ultrathin-layer analytical isoeletcric focusing electrophorsi s, the β-lactamase focusing patterns of each strain (9 Ceftazidime-resistant strains, in which 7 E.cloacae, 2 P. aeruginosa) consisted of a single band except that enzyme R27853 had 2 bands. The isoelectric points (pI) of β-lactamases R10C6, R8701 and R884 were the same as that of standard chromosamal-mediated β-lactamase D-31 (pI = 8.6), the pis of β-lactamases R222 and R1029 were similar to that of standard chromosamal-mediated β-lactamase P-99 (pI = 8.0). In addition,β-lactamase R27853 (pI = 8.2) and β-lactamases R471, R585 and R875(pI = 8.4)respetcively may be new chromosamal β-lacta mases. The experiments suggested that the ultrathin-layer isoelectric focusing electrophore-sis characterized by its high resolution, high speed, versatility and simplicity of operation, and recommends itself as a good analytical technique for research study and routine applications.

关 键 词：内酰胺酶 超薄层 等电聚焦 

分 类 号：R927.1[医药卫生—药学]