氨基糖苷磷酸转移酶的显色检测
Chromogenic detection of aminoglycoside phosphotransferases.
作者信息
Amoroso A M, Gutkind G O
机构信息
Cátedra de Microbiología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Argentina.
出版信息
Antimicrob Agents Chemother. 1998 Feb;42(2):228-30. doi: 10.1128/AAC.42.2.228.
A coupled chromogenic reaction (based on an agar overlay combining NADH, pyruvate kinase, lactate dehydrogenase, phosphoenolpyruvate, ATP, and kanamycin sulfate with thiazolyl blue-phenazine methosulfate for detection of NADH consumption) was optimized for the detection of aminoglycoside phosphotransferases (APHs). When used after analytical isoelectrofocusing of bacterial extracts from APH-producing strains, this method revealed one band in each of two strains with a genetically confirmed APH (3') I and two bands in another strain with both APH (3') I and APH (3') VI, whereas no bands were detected in susceptible control strains or in aminoglycoside-resistant microorganisms without APH genes.
一种偶联显色反应（基于将烟酰胺腺嘌呤二核苷酸（NADH）、丙酮酸激酶、乳酸脱氢酶、磷酸烯醇丙酮酸、三磷酸腺苷（ATP）和硫酸卡那霉素与噻唑蓝-吩嗪硫酸甲酯相结合的琼脂覆盖物用于检测NADH消耗）被优化用于检测氨基糖苷磷酸转移酶（APHs）。当用于对产生APH的菌株的细菌提取物进行分析等电聚焦后时，该方法在两个经基因确认具有APH(3')I的菌株中各显示一条带，在另一个同时具有APH(3')I和APH(3')VI的菌株中显示两条带，而在敏感对照菌株或无APH基因的氨基糖苷抗性微生物中未检测到条带。
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