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利用 RESIST-3 O.K.N. 免疫检测法直接从增菌血培养物中快速检测巴西的主要碳青霉烯酶。

Rapid detection of the main carbapenemases in Brazil directly from spiked blood culture using the RESIST-3 O.K.N. immunoassay.

机构信息

LABRESIS (Laboratório de Pesquisa em Resistência Bacteriana), Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos 2350, Porto Alegre, RS, 90035-903, Brazil.

Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.

出版信息

Braz J Microbiol. 2019 Jul;50(3):657-662. doi: 10.1007/s42770-019-00109-y. Epub 2019 Jul 4.

Abstract

The emergence of carbapenem-resistant Enterobacterales (CRE) is a matter of public health concern. Carbapenemases are the main mechanism of resistance among CRE, and its rapid detection is essential. The detection of carbapenemases usually requires culture-based methods and molecular assays, which may be costly and need long turnaround times. Recently, an easy and rapid immunochromatographic assay for carbapenemases (OXA-48, KPC, and NDM) detection based in lateral flow immunoassay with specific monoclonal antibodies on a nitrocellulose membrane has been developed. We aimed to evaluate the RESIST-3 O.K.N. in colonies from pure culture as well as in spiked blood cultures with Enterobacterales. All carbapenemase producers (CP) presenting the OXA-48-like, KPC, and NDM enzymes presented positive results in both pure colonies and spiked blood cultures. None of the carbapenemase non-producers (CNP) presented positive results in the tests. A total of 97% CP isolates presented positive results in pure colonies in less than 5 min. For CP directly from blood culture, the mean time to positivity for OXA-48-like and KPC was 1 min, whereas it was 25 min for NDM. Our results indicate that this immunoassay can be used to detect carbapenemases directly from blood culture bottles in a routine diagnostic laboratory, which would reduce the turnaround time of CP detection.

摘要

碳青霉烯类耐药肠杆菌科 (CRE) 的出现是一个公共卫生关注的问题。碳青霉烯酶是 CRE 耐药的主要机制,其快速检测至关重要。碳青霉烯酶的检测通常需要基于培养的方法和分子检测,这可能成本高昂且需要较长的周转时间。最近,已经开发出一种基于侧向流动免疫分析的简便快速免疫层析测定法,用于检测碳青霉烯酶 (OXA-48、KPC 和 NDM),该方法在硝酸纤维素膜上使用特定的单克隆抗体。我们旨在评估 RESIST-3 O.K.N. 在纯培养物的菌落以及添加肠杆菌科的血培养物中的检测效果。所有产碳青霉烯酶的细菌 (CP) ,其 OXA-48 样、KPC 和 NDM 酶均在纯菌落和添加血培养物中呈现阳性结果。没有碳青霉烯酶非生产者 (CNP) 的测试呈阳性结果。在不到 5 分钟的时间内,97%的 CP 分离株在纯菌落中呈阳性结果。对于直接来自血培养物的 CP,OXA-48 样和 KPC 的阳性时间平均为 1 分钟,而 NDM 的阳性时间为 25 分钟。我们的结果表明,这种免疫测定法可用于常规诊断实验室直接从血培养瓶中检测碳青霉烯酶,从而缩短 CP 检测的周转时间。

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