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利用表面多孔液相色谱-串联质谱法快速灵敏地检测鲍曼不动杆菌中的碳青霉烯酶活性。

Rapid and sensitive detection of carbapenemase activity in Acinetobacter baumannii using superficially porous liquid chromatography-tandem mass spectrometry.

机构信息

Department of Laboratory Medicine and Biotechnology, Tzu Chi University, Hualien, Taiwan.

Department of Laboratory Medicine and Biotechnology, Tzu Chi University, Hualien, Taiwan; Department of Laboratory Medicine, Buddhist Tzu Chi General Hospital, Hualien, Taiwan.

出版信息

J Microbiol Immunol Infect. 2016 Dec;49(6):910-917. doi: 10.1016/j.jmii.2015.08.001. Epub 2015 Aug 14.

Abstract

BACKGROUND

The emergence and spread of carbapenem-resistant Acinetobacter baumannii poses a challenge for optimizing antibiotic therapies and preventing outbreaks. Traditional phenotypic assays such as the modified Hodge test (MHT) or polymerase chain reaction (PCR)-based detection of the carbapenemase genes are time-consuming and complicated. Therefore, new approaches for the efficient detection of carbapenemase-producing A. baumannii are urgently required.

METHODS

In this study, we used the superficially porous liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay to measure carbapenem hydrolysis in a solution spiked with test strains of A. baumannii. The rate of carbapenem hydrolysis during incubation was expressed as the ratio of the carbapenem peak area of the test A. baumannii strains to the noncarbapenemase-producing A. baumannii ATCC 17978. This method can accurately measure the carbapenem hydrolysis rate and, therefore, can effectively identify carbapenemase-producing strains within 75 minutes.

RESULTS

A total of 112 A. baumannii strains were used in this study, including 103 clinical isolates with 68 carbapenem-resistant strains and 35 carbapenem-susceptible strains, seven ATCC strains and two selected mutants. The results of the superficially porous LC-MS/MS assay showed higher detection sensitivity compared to the results of the MHT.

CONCLUSION

Our results demonstrate the ability of the former method to routinely detect carbapenemase-producing A. baumannii.

摘要

背景

耐碳青霉烯鲍曼不动杆菌的出现和传播对优化抗生素治疗和预防暴发提出了挑战。传统的表型检测方法,如改良 Hodge 试验(MHT)或基于聚合酶链反应(PCR)的碳青霉烯酶基因检测,既费时又复杂。因此,迫切需要新的方法来有效检测产碳青霉烯酶的鲍曼不动杆菌。

方法

在这项研究中,我们使用表面多孔液相色谱-串联质谱(LC-MS/MS)测定法来测量含测试鲍曼不动杆菌菌株的溶液中碳青霉烯的水解。孵育过程中碳青霉烯水解的速度表示为测试鲍曼不动杆菌菌株的碳青霉烯峰面积与非产碳青霉烯酶鲍曼不动杆菌 ATCC 17978 的比值。该方法可准确测量碳青霉烯水解率,因此可在 75 分钟内有效识别产碳青霉烯酶菌株。

结果

本研究共使用了 112 株鲍曼不动杆菌菌株,包括 103 株临床分离株,其中 68 株为耐碳青霉烯菌株,35 株为碳青霉烯敏感菌株,7 株 ATCC 菌株和 2 株选定的突变株。表面多孔 LC-MS/MS 测定法的结果显示出比 MHT 更高的检测灵敏度。

结论

我们的结果表明,该方法能够常规检测产碳青霉烯酶的鲍曼不动杆菌。

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