Modified Carba PBP test for rapid detection and differentiation between different classes of carbapenemases in Enterobacterales

IF 5.3 2区化学 Q1 CHEMISTRY, ANALYTICAL Microchimica Acta Pub Date : 2024-12-05 DOI:10.1007/s00604-024-06859-3

Xiaonan Wang, Zhimin Lu, Leina Dou, Licai Ma, Tong He, Chenxi Gao, Xiangjun Zhao, Jin Tao, Liang Luo, Qing Li, Yang Wang, Yingbo Shen, Jianzhong Shen, Zhanhui Wang, Kai Wen

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Abstract

An advanced biochemical assay named modified Carba PBP test was innovated to identify and differentiate distinct categories of clinically significant carbapenemases (Ambler classes A, B, and D) within the Enterobacterales. The mechanism of mCarba PBP hinges on two core attributes: (i) the hydrolysis of the meropenem substrate by various carbapenemases, (ii) the immobilized penicillin and free meropenem in their affinity to interact with a limited quantity of penicillin-binding protein (PBP). Specific inhibitors for class A (phenylboronic acid, PBA) and class B (ethylenediaminetetraacetic acid, EDTA) were employed to inhibit the hydrolysis activity of carbapenemase and facilitate the classification of carbapenemase classes within 25 min. A comprehensive evaluation was undertaken using 94 clinical Enterobacterales isolates, comprising 75 carbapenemase-producing strains and 19 non-carbapenemase-producing strains. Its overall specificity and sensitivity were 100% and 97.3%, respectively, including detection of all types of OXA-48-like carbapenemases. For precise carbapenemase type identification, the assay exhibited remarkable sensitivities for class A, class B, and class D detection at 94.7%, 100%, and 100%, respectively. This user-friendly test presents a promising tool for carbapenemase identification, refining the selection of β-lactam/β-endoenzyme inhibitor combinations for effectively treating infections due to carbapenemase-producing organisms.

Graphical Abstract

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改良的Carba PBP法在肠杆菌中不同种类碳青霉烯酶的快速检测和区分

一种名为改良Carba PBP测试的先进生化试验被创新用于识别和区分肠杆菌中不同类别的临床重要碳青霉烯酶（Ambler类A， B和D）。mCarba PBP的机制取决于两个核心属性：(i)各种碳青霉烯酶水解美罗培南底物，（ii）固定化青霉素和游离美罗培南与有限数量的青霉素结合蛋白（PBP）相互作用的亲和力。采用A类（苯硼酸，PBA）和B类（乙二胺四乙酸，EDTA）特异性抑制剂抑制碳青霉烯酶的水解活性，并在25 min内对碳青霉烯酶进行分类。采用94株临床肠杆菌进行综合评价，其中75株产碳青霉烯酶菌株和19株非产碳青霉烯酶菌株。其总体特异性和敏感性分别为100%和97.3%，包括检测所有类型的oxa -48样碳青霉烯酶。对于碳青霉烯酶类型的精确鉴定，该方法对A类、B类和D类的检测灵敏度分别为94.7%、100%和100%。这种用户友好的测试为碳青霉烯酶鉴定提供了一个很有前途的工具，改进了β-内酰胺/β-内酶抑制剂组合的选择，有效地治疗碳青霉烯酶产生生物引起的感染。图形抽象

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来源期刊

Microchimica Acta 化学-分析化学

CiteScore

9.80

自引率

5.30%

发文量

410

审稿时长

2.7 months

期刊介绍： As a peer-reviewed journal for analytical sciences and technologies on the micro- and nanoscale, Microchimica Acta has established itself as a premier forum for truly novel approaches in chemical and biochemical analysis. Coverage includes methods and devices that provide expedient solutions to the most contemporary demands in this area. Examples are point-of-care technologies, wearable (bio)sensors, in-vivo-monitoring, micro/nanomotors and materials based on synthetic biology as well as biomedical imaging and targeting.