Wang Xiaonan, Lu Zhimin, Dou Leina, Ma Licai, He Tong, Gao Chenxi, Zhao Xiangjun, Tao Jin, Luo Liang, Li Qing, Wang Yang, Shen Yingbo, Shen Jianzhong, Wang Zhanhui, Wen Kai
National Key Laboratory of Veterinary Public Health and Safety, College of Veterinary Medicine, China Agricultural University, Yuanmingyuan West Road 2#, Haidian District, Beijing, 100193, People's Republic of China.
College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, Shaanxi, People's Republic of China.
Mikrochim Acta. 2024 Dec 5;192(1):7. doi: 10.1007/s00604-024-06859-3.
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.
一种名为改良碳青霉烯类青霉素结合蛋白（Carba PBP）试验的先进生化检测方法被创新出来，用于鉴定和区分肠杆菌科细菌中临床上具有重要意义的不同类别碳青霉烯酶（安布勒A、B和D类）。改良碳青霉烯类青霉素结合蛋白（mCarba PBP）试验的机制取决于两个核心特性：（i）各种碳青霉烯酶对美罗培南底物的水解作用；（ii）固定化青霉素和美罗培南与有限量青霉素结合蛋白（PBP）相互作用的亲和力。使用A类（苯硼酸，PBA）和B类（乙二胺四乙酸，EDTA）的特异性抑制剂来抑制碳青霉烯酶的水解活性，并在25分钟内促进碳青霉烯酶类别的分类。使用94株临床肠杆菌科分离株进行了全面评估，其中包括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。对于精确的碳青霉烯酶类型鉴定，该检测方法对A类、B类和D类检测的敏感性分别为94.7%、100%和100%。这种用户友好的检测方法为碳青霉烯酶鉴定提供了一种有前景的工具，有助于优化β-内酰胺/β-内酰胺酶抑制剂组合的选择，以有效治疗由产碳青霉烯酶生物体引起的感染。
