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评估与血流感染相关的耐碳青霉烯类分离株中 NG-Test CARBA 5 多重免疫层析法和 Cepheid Xpert CARBA-R 检测的应用。

Evaluating NG-Test CARBA 5 Multiplex Immunochromatographic and Cepheid Xpert CARBA-R Assays among Carbapenem-Resistant Isolates Associated with Bloodstream Infection.

机构信息

Department of Laboratory Medicine, National Taiwan University Hospitalgrid.412094.a, National Taiwan University College of Medicine, Taipei, Taiwan.

Department of Internal Medicine, National Taiwan University Hospitalgrid.412094.a, National Taiwan University College of Medicine, Taipei, Taiwan.

出版信息

Microbiol Spectr. 2022 Feb 23;10(1):e0172821. doi: 10.1128/spectrum.01728-21. Epub 2022 Jan 12.

Abstract

Decreased susceptibility to carbapenems in is an emerging concern. Conventional methods with short turnaround times are crucial for therapeutic decisions and infection control. In the current study, we used the Xpert CARBA-R (Cepheid, Sunnyvale, CA, USA) and the NG-Test CARBA 5 (NG Biotech, Guipry, France) assays for carbapenemase detection in 214 carbapenem-resistant (CRE) blood isolates. We used the modified carbapenem inactivation method, conventional PCR, and sequencing to determine the production of five common carbapenemase families and their subtypes. We performed -genotyping for all CR-Klebsiella pneumoniae (CRKP) and multilocus sequence typing for all carbapenemase-producing CRE isolates to reveal their genetic relatedness. The results showed a sensitivity of 99.8% and a specificity of 100% by the Xpert assay, and a sensitivity of 100% and a specificity of 99% by the NG-Test in detecting carbapenemases of 84 CRKP isolates with only one (VIM-1+IMP-8) failure in both tests. For CR-Escherichia coli, four carbapenemase-producing isolates were detected accurately for their subtypes. The two major clones of carbapenemase-producing CRKP isolates in Taiwan were ST11-K47 producing KPC-2 (= 47) and ST11-K64 producing OXA-48-like (= 9). Our results support the use of either test in routine laboratories for the rapid detection of common carbapenemases. Caution should be taken using the Xpert assay in areas with a high prevalence of CRE carrying . Carbapenemase-producing (CPE) are emerging worldwide, causing nosocomial outbreaks and even community-acquired infections since their appearance 2 decades ago. Our previous national surveillance of CPE isolates in Taiwan identified five carbapenemase families (KPC, OXA, NDM, VIM, and IMP) with the KPC-2 and OXA-48-like types predominant. Timely detection and classification of carbapenemases in CPE may be a useful test to guide optimal therapy and infection control. Genetic detection methods using the Xpert CARBA-R assay and the immunochromatographic assay using the NG-Test CARBA 5 have been validated with the advantage of short turnaround time. Our study demonstrated that the NG and Xpert assays are convenient methods to accurately identify carbapenemases in carbapenem-resistant Klebsiella pneumoniae and carbapenem-resistant Escherichia coli blood isolates. Detecting IMP variants remains challenging, and the results of Xpert CARBA-R assay should be carefully interpreted.

摘要

碳青霉烯类药物耐药肠杆菌科细菌(CRE)的易感性降低是一个新出现的问题。具有较短周转时间的常规方法对于治疗决策和感染控制至关重要。在本研究中,我们使用了 Xpert CARBA-R(Cepheid,加利福尼亚州桑尼维尔)和 NG-Test CARBA 5(NG Biotech,法国吉普鲁)检测 214 株碳青霉烯类耐药肠杆菌科细菌(CRE)血分离株中的碳青霉烯酶。我们使用改良的碳青霉烯类药物灭活法、常规 PCR 和测序来确定五种常见的碳青霉烯酶家族及其亚型的产生。我们对所有产碳青霉烯酶肺炎克雷伯菌(CRKP)进行了 - 基因分型,对所有产碳青霉烯酶 CRE 分离株进行了多位点序列分型,以揭示它们的遗传关系。结果显示,Xpert 检测法的敏感性为 99.8%,特异性为 100%,NG-Test 检测法的敏感性为 100%,特异性为 99%,在检测仅 1 个(VIM-1+IMP-8)失败的 84 株 CRKP 分离株中的碳青霉烯酶时,两种检测方法均表现出良好的效果。对于产碳青霉烯酶大肠埃希菌,4 株产碳青霉烯酶分离株的亚型检测准确。台湾产碳青霉烯酶 CRKP 分离株的两个主要克隆是 ST11-K47 产 KPC-2(=47)和 ST11-K64 产 OXA-48 样(=9)。我们的结果支持在常规实验室中使用这两种检测方法快速检测常见的碳青霉烯酶。在存在高流行率携带碳青霉烯酶的肠杆菌科细菌(CRE)的地区,使用 Xpert 检测法应谨慎。自 20 年前出现以来,产碳青霉烯酶肠杆菌科细菌(CPE)在全球范围内不断出现,导致医院感染甚至社区获得性感染。我们之前在台湾进行的全国性 CPE 分离株监测确定了五种碳青霉烯酶家族(KPC、OXA、NDM、VIM 和 IMP),其中 KPC-2 和 OXA-48 样型占主导地位。及时检测和分类 CPE 中的碳青霉烯酶可能是指导最佳治疗和感染控制的有用检测方法。使用 Xpert CARBA-R 检测法的基因检测方法和使用 NG-Test CARBA 5 的免疫层析检测法已经过验证,具有周转时间短的优势。我们的研究表明,NG 和 Xpert 检测法是准确鉴定碳青霉烯类耐药肺炎克雷伯菌和碳青霉烯类耐药大肠埃希菌血分离株中碳青霉烯酶的便捷方法。检测 IMP 变体仍然具有挑战性,Xpert CARBA-R 检测法的结果应仔细解释。

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