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镧系螯合物标记的侧向流动分析用于青霉素结合蛋白快速多重检测β-内酰胺类抗生素。

Europium chelate-labeled lateral flow assay for rapid and multiple detection of β-lactam antibiotics by the penicillin-binding protein.

机构信息

Medical Science Research Center, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, 100730, China.

出版信息

Anal Methods. 2020 Jul 28;12(28):3645-3653. doi: 10.1039/d0ay01140a. Epub 2020 Jul 6.

Abstract

For the multiple and rapid detection of β-lactams, the broadly specific penicillin-binding protein (PBP) that recognizes the β-lactam structure was prepared. A chromatographic strip based on europium chelate-loaded fluorescent microspheres was assembled with a goat anti-mouse antibody (C line) and penicillin coating (T line). The penicillin coating competes with free β-lactam antibiotics to bind PBP-labeled fluorescent microspheres. The strip can theoretically detect all kinds of β-lactams, including amoxicillin, ampicillin, oxacillin, ceftazidime, lenampicillin, cefoperazone, sultamicillin, cefotaxime, ceftriaxone, sulbenicillin, piperacillin, cephalothin, flucloxacillin, and mezlocillin, in samples within 10 min. The europium chelate-labeled lateral flow assay does not cross-react with other antibiotics, including chloramphenicol, tetracycline, sulfamethazine, enrofloxacin, gentamicin or lincomycin. In short, we developed a very useful method for preliminary screening of β-lactams.

摘要

为了实现β-内酰胺类抗生素的多重快速检测,我们制备了一种广谱特异性的青霉素结合蛋白(PBP),该蛋白能够识别β-内酰胺结构。我们将负载铕螯合物的荧光微球组装成一种基于色谱的检测条,其中包含山羊抗小鼠抗体(C 线)和青霉素涂层(T 线)。青霉素涂层与游离的β-内酰胺类抗生素竞争,与 PBP 标记的荧光微球结合。该检测条理论上可以在 10 分钟内检测到样品中的所有β-内酰胺类抗生素,包括阿莫西林、氨苄西林、苯唑西林、头孢他啶、仑氨西林、头孢哌酮、舒巴坦、头孢噻肟、头孢曲松、舒巴坦、哌拉西林、头孢噻吩、氟氯西林和头孢美唑。铕螯合物标记的侧向流动检测法不会与其他抗生素(如氯霉素、四环素、磺胺嘧啶、恩诺沙星、庆大霉素或林可霉素)发生交叉反应。总之,我们开发了一种非常有用的β-内酰胺类抗生素初步筛选方法。

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