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评价一种用于检测产 GES 的革兰氏阴性细菌的新型快速免疫层析检测法。

Evaluation of a new rapid immunochromatographic assay for the detection of GES-producing Gram-negative bacteria.

机构信息

Team 'Resist', UMR1184 'Immunology of Viral, Auto-Immune, Hematological and Bacterial diseases (IMVA-HB),' INSERM, Université Paris-Saclay, CEA, LabEx LERMIT, Faculty of Medicine, Le Kremlin-Bicêtre, France.

Bacteriology-Hygiene Unit, Assistance Publique-Hôpitaux de Paris, AP-HP Paris-Saclay, Bicêtre Hospital, Le Kremlin-Bicêtre, France.

出版信息

J Antimicrob Chemother. 2023 May 3;78(5):1282-1287. doi: 10.1093/jac/dkad090.

Abstract

BACKGROUND

As carbapenemase-producing Enterobacterales are increasingly reported worldwide, their rapid detection is crucial to reduce their spread and prevent infections and outbreaks. Lateral flow immunoassays (LFIAs) have become major tools for the detection of carbapenemases. However, as for most commercially available assays, only the five main carbapenemases are targeted.

OBJECTIVES

Here, we have developed and evaluated an LFIA prototype for the rapid and reliable detection of the increasingly identified GES-type β-lactamases.

METHODS

The GES LFIA was validated on 103 well-characterized Gram-negative isolates expressing various β-lactamases grown on Mueller-Hinton (MH) agar, chromogenic, and chromogenic/selective media.

RESULTS

The limit of detection of the assay was 106 cfu per test with bacteria grown on MH agar plates. GES LFIA accurately detected GES-type β-lactamases irrespective of the culture media and the bacterial host. The GES LFIA was not able to distinguish between GES-ESBLs and GES-carbapenemases. Because GES enzymes are still rare, their detection as an ESBL or a carbapenemase remains important, especially because extensive use of carbapenems to treat ESBL infections may select for GES variants capable of hydrolysing carbapenems.

CONCLUSIONS

The GES LFIA is efficient, rapid and easy to implement in the routine workflow of a clinical microbiology laboratory for the confirmation of GES-type β-lactamases. Combining it with immunochromatographic assays targeting the five main carbapenemases (KPC, NDM, VIM, IMP and OXA-48) would improve the overall sensitivity for the most frequently encountered carbapenemases and ESBLs, especially in non-fermenters.

摘要

背景

随着产碳青霉烯酶肠杆菌科细菌在全球范围内的报道越来越多,快速检测对于减少其传播、预防感染和爆发至关重要。侧向流动免疫分析(LFIAs)已成为检测碳青霉烯酶的主要工具。然而,与大多数市售检测方法一样,这些方法仅针对五种主要的碳青霉烯酶。

目的

本研究开发并评估了一种用于快速可靠检测日益增多的 GES 型β-内酰胺酶的 LFIA 原型。

方法

在 Mueller-Hinton(MH)琼脂、显色和显色/选择性培养基上生长的 103 种表达各种β-内酰胺酶的经过充分特征鉴定的革兰氏阴性分离株上验证了 GES LFIA。

结果

该检测方法的检测限为在 MH 琼脂平板上培养的细菌每测试 106 cfu。GES LFIA 可准确检测 GES 型β-内酰胺酶,无论培养介质和细菌宿主如何。GES LFIA 无法区分 GES-ESBLs 和 GES-碳青霉烯酶。由于 GES 酶仍然很少见,因此将其作为 ESBL 或碳青霉烯酶进行检测仍然很重要,尤其是因为广泛使用碳青霉烯类药物治疗 ESBL 感染可能会选择能够水解碳青霉烯类药物的 GES 变体。

结论

GES LFIA 快速、高效且易于在临床微生物学实验室的常规工作流程中实施,用于确认 GES 型β-内酰胺酶。将其与针对五种主要碳青霉烯酶(KPC、NDM、VIM、IMP 和 OXA-48)的免疫层析检测方法相结合,将提高最常见的碳青霉烯酶和 ESBL 的总体敏感性,尤其是在非发酵菌中。

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