Rapid detection of plasmid-mediated AmpC-producers by eazyplex® SuperBug AmpC assay compared to whole-genome sequencing

IF 1.7 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Journal of microbiological methods Pub Date : 2024-04-19 DOI:10.1016/j.mimet.2024.106938

Vladimira Hinić , Helena M.B. Seth-Smith , Sabrina Stammler , Adrian Egli

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引用次数: 0

Abstract

Current methods for plasmid-mediated AmpC β-lactamase (pAmpC) detection in routine microbiological laboratories are based on various phenotypic tests. Eazyplex®SuperBug AmpC assay is a molecular assay based on isothermal amplification for rapid detection of the most common pAmpC types from bacterial culture: CMY-2 group, DHA, ACC and MOX. Our aim was to evaluate the diagnostic performance of this assay. The assay was evaluated on 64 clinical isolates of Enterobacterales without chromosomal inducible AmpC, and with phenotypically confirmed AmpC production. The results were confirmed, and isolates further characterized by whole-genome sequencing (WGS). eazyplex®SuperBug AmpC assay correctly detected the two most common pAmpC types CMY-2 group (16/16) and DHA (19/19). Detection of ACC and MOX could not be evaluated on our set of isolates since there was only one isolate harbouring ACC and none with MOX. pAmpC encoding genes could be detected in only eight of 36 investigated Escherichia coli isolates. The remaining 28 E. coli isolates harboured previously described mutations in the bla_EC promoter, leading to the overexpression of chromosomally encoded E. coli specific AmpC β-lactamase. All results were 100% concordant with the results of WGS. eazyplex®SuperBug AmpC assay enabled rapid and reliable detection of pAmpC-encoding genes in Enterobacterales like Klebsiella spp. and Proteus spp. and the distinction between plasmid-mediated and chromosomally encoded AmpC in E. coli.

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与全基因组测序法相比，eazyplex® SuperBug AmpC 检测法可快速检测质粒介导的 AmpC 生产者

目前常规微生物实验室检测质粒介导的 AmpC β-内酰胺酶（pAmpC）的方法主要基于各种表型检测。Eazyplex®SuperBug AmpC 检测试剂盒是一种基于等温扩增的分子检测试剂盒，可快速检测细菌培养物中最常见的 pAmpC 类型：CMY-2 组、DHA、ACC 和 MOX。我们的目的是评估该检测方法的诊断性能。我们对 64 个不含染色体诱导型 AmpC 的肠杆菌临床分离物进行了评估，这些分离物具有经表型证实的 AmpC 生产。eazyplex®SuperBug AmpC 检测法正确检测出两种最常见的 pAmpC 类型 CMY-2 组（16/16）和 DHA（19/19）。由于只有一个分离物携带 ACC，没有一个分离物携带 MOX，因此无法对我们的分离物进行 ACC 和 MOX 的检测评估。在调查的 36 个大肠埃希菌分离物中，只有 8 个分离物能检测到 pAmpC 编码基因。其余 28 个大肠杆菌分离物的 blaEC 启动子中含有先前描述过的突变，导致染色体编码的大肠杆菌特异性 AmpC β-内酰胺酶过度表达。eazyplex®SuperBug AmpC 检测法能够快速、可靠地检测克雷伯氏菌属和变形杆菌属等肠杆菌科细菌的 pAmpC 编码基因，并区分大肠杆菌中质粒介导的 AmpC 和染色体编码的 AmpC。

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来源期刊

Journal of microbiological methods 生物-生化研究方法

CiteScore

4.30

自引率

4.50%

发文量

151

审稿时长

29 days

期刊介绍： The Journal of Microbiological Methods publishes scholarly and original articles, notes and review articles. These articles must include novel and/or state-of-the-art methods, or significant improvements to existing methods. Novel and innovative applications of current methods that are validated and useful will also be published. JMM strives for scholarship, innovation and excellence. This demands scientific rigour, the best available methods and technologies, correctly replicated experiments/tests, the inclusion of proper controls, calibrations, and the correct statistical analysis. The presentation of the data must support the interpretation of the method/approach. All aspects of microbiology are covered, except virology. These include agricultural microbiology, applied and environmental microbiology, bioassays, bioinformatics, biotechnology, biochemical microbiology, clinical microbiology, diagnostics, food monitoring and quality control microbiology, microbial genetics and genomics, geomicrobiology, microbiome methods regardless of habitat, high through-put sequencing methods and analysis, microbial pathogenesis and host responses, metabolomics, metagenomics, metaproteomics, microbial ecology and diversity, microbial physiology, microbial ultra-structure, microscopic and imaging methods, molecular microbiology, mycology, novel mathematical microbiology and modelling, parasitology, plant-microbe interactions, protein markers/profiles, proteomics, pyrosequencing, public health microbiology, radioisotopes applied to microbiology, robotics applied to microbiological methods,rumen microbiology, microbiological methods for space missions and extreme environments, sampling methods and samplers, soil and sediment microbiology, transcriptomics, veterinary microbiology, sero-diagnostics and typing/identification.