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一种基于聚对二甲苯基质芯片的激光解吸/电离质谱法的高灵敏度碳青霉烯酶检测方法。

A highly sensitive carbapenemase assay using laser desorption/ionization mass spectrometry based on a parylene-matrix chip.

作者信息

Park Jong-Min, Kim Jo-Il, Noh Joo-Yoon, Kim Mira, Kang Min-Jung, Pyun Jae-Chul

机构信息

Department of Materials Science and Engineering, Yonsei University, 134 Shinchon-dong, Seodaemun-gu Seoul, 03722, Korea.

Korea Institute of Science and Technology (KIST), Hwarang-ro 14-gil 5, Seongbuk-gu, Seoul, 02792, Korea.

出版信息

Enzyme Microb Technol. 2017 Sep;104:56-68. doi: 10.1016/j.enzmictec.2017.05.010. Epub 2017 Jun 7.

Abstract

A quantitative carbapenemase assay was developed using laser desorption/ionization mass spectrometry (LDI-MS) based on a parylene-matrix chip. As a first step, the reproducibility (spot-to-spot, shot-to-shot, and day-to-day) of LDI-MS based on a parylene-matrix chip and the quantification ranges for four carbapenem antibiotics (doripenem, ertapenem, imipenem, and meropenem) were determined. A carbapenem-susceptibility test was performed using the four carbapenems and 51 bacterial strains that displayed (1) carbapenem resistance with carbapenemase, (2) carbapenem resistance without carbapenemase, or (3) carbapenem susceptibility. The susceptibility test results showed that LDI-MS based on a parylene-matrix chip was more sensitive and selective for detecting the carbapenemase reaction than conventional MALDI-TOF MS based on a 2,5-dihydroxybenzoic acid matrix.

摘要

基于聚对二甲苯基质芯片,利用激光解吸/电离质谱(LDI-MS)开发了一种碳青霉烯酶定量检测方法。第一步,确定了基于聚对二甲苯基质芯片的LDI-MS的重现性(点与点、次与次以及日复一日)以及四种碳青霉烯类抗生素(多利培南、厄他培南、亚胺培南和美罗培南)的定量范围。使用这四种碳青霉烯类抗生素对51株细菌进行了碳青霉烯敏感性试验,这些细菌表现为:(1)具有碳青霉烯酶的碳青霉烯耐药性;(2)无碳青霉烯酶的碳青霉烯耐药性;或(3)碳青霉烯敏感性。敏感性试验结果表明,基于聚对二甲苯基质芯片的LDI-MS在检测碳青霉烯酶反应方面比基于2,5-二羟基苯甲酸基质的传统基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)更灵敏、更具选择性。

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