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A Multiplex Lateral Flow Immunoassay for the Rapid Identification of NDM-, KPC-, IMP- and VIM-type and OXA-48-like Carbapenemase-Producing Enterobacteriaceae

Journal of Antimicrobial Chemotherapy(2017)SCI 2区SCI 1区

Univ Paris Saclay

Cited 168|Views26
Abstract
Abstract Objectives The global spread of carbapenemase-producing Enterobacteriaceae represents a substantial challenge in clinical practice and rapid and reliable detection of these organisms is essential. The aim of this study was to develop and validate a lateral flow immunoassay (Carba5) for the detection of the five main carbapenemases (KPC-, NDM-, VIM- and IMP-type and OXA-48-like). Methods Carba5 was retrospectively and prospectively evaluated using 296 enterobacterial isolates from agar culture. An isolated colony was suspended in extraction buffer and then loaded on the manufactured Carba5. Results All 185 isolates expressing a carbapenemase related to one of the Carba5 targets were correctly and unambiguously detected in <15 min. All other isolates gave negative results except those producing OXA-163 and OXA-405, which are considered low-activity carbapenemases. No cross-reaction was observed with non-targeted carbapenemases, ESBLs, AmpCs or oxacillinases (OXA-1, -2, -9 and -10). Overall, this assay reached 100% sensitivity and 95.3% (retrospectively) to 100% (prospectively) specificity. Conclusions Carba5 is efficient, rapid and easy to implement in the routine workflow of a clinical microbiology laboratory for confirmation of the five main carbapenemases encountered in Enterobacteriaceae.
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要点】:本文研发并验证了一种名为Carba5的侧流免疫分析技术,用于快速检测五种主要的碳青霉烯酶(KPC-、NDM-、VIM-、IMP-型及OXA-48样酶),具有高效性、快速性和易操作的特点。

方法】:通过将分离的菌落悬浮于提取缓冲液中,随后加载到Carba5侧流免疫分析装置上进行检测。

实验】:实验使用296株肠杆菌属分离株,包括回顾性和前瞻性评估,结果显示Carba5在15分钟内准确无误地检测出所有表达与目标相关的碳青霉烯酶的185株菌,总体达到100%的灵敏度和95.3%(回顾性)至100%(前瞻性)的特异性。