Faculty of Medicine and Nursing, Immunology, Microbiology and Parasitology, University of the Basque Country UPV/EHU, Bilbao, Spain.
German Center for Infection Research (DZIF), partner site Bonn-Cologne, Cologne, Germany.
J Med Microbiol. 2021 Mar;70(3). doi: 10.1099/jmm.0.001310. Epub 2021 Jan 15.
Gram-negative bacteria are a common source of infection both in hospitals and in the community, and antimicrobial resistance is frequent among them, making antibiotic therapy difficult, especially when these isolates carry carbapenem resistance determinants.. A simple method to detect all the commonly found carbapenemases in Germany was not available. The aim of this study was to develop a multiplex PCR for the rapid and reliable identification of the most prevalent carbapenemase-encoding genes in Gram-negative bacteria in Germany. Data from the German Gram-negative reference laboratory revealed the most prevalent carbapenemase groups in Germany were (in order of prevalence): , , , , , , , , , , IS , , and . We developed and tested two multiplex PCRs against 83 carbapenem-resistant Gram-negative clinical isolates. Primers were designed for each carbapenemase group within conserved regions of the encoding genes obtained from publicly available databases. Multiplex-1 included the carbapenemase groups , , , , and , while multiplex-2 included , , , , IS and . In the initial evaluation, all but one of the carbapenemases encoded by 75 carbapenemase-positive isolates were detected using the two multiplex PCRs, while no false-positive results were obtained from the remaining eight isolates. After evaluation, we tested 546 carbapenem-resistant isolates using the multiplex PCRs, and all carbapenemases were detected. A rapid and reliable method was developed for detection and differentiation of 12 of the most prevalent carbapenemase groups found in Germany. This method allows for the rapid testing of clinical isolates prior to species identification and does not require prior phenotypical characterization, constituting a rapid and valuable tool in the management of infections in hospitals.
革兰氏阴性菌是医院和社区感染的常见来源，它们普遍存在抗生素耐药性，使抗生素治疗变得困难，尤其是当这些分离株携带碳青霉烯类耐药决定因素时。德国没有一种简单的方法可以检测所有常见的碳青霉烯酶。本研究旨在开发一种多重 PCR 方法，用于快速可靠地鉴定德国革兰氏阴性菌中最常见的碳青霉烯酶编码基因。来自德国革兰氏阴性参考实验室的数据显示，德国最常见的碳青霉烯酶组依次为： 、 、 、 、 、 、 、 、 IS 、 、 和 。我们针对 83 株耐碳青霉烯的革兰氏阴性临床分离株开发并测试了两种多重 PCR。引物是根据从公共数据库中获得的编码基因的保守区域设计的，用于每个碳青霉烯酶组。多重 PCR-1 包括 、 、 、 、 和 ，而多重 PCR-2 包括 、 、 、 IS 和 。在最初的评估中，使用两种多重 PCR 检测到 75 株碳青霉烯酶阳性分离株中编码的除一种碳青霉烯酶以外的所有碳青霉烯酶，而其余 8 株分离株未得到假阳性结果。经过评估，我们使用多重 PCR 测试了 546 株耐碳青霉烯的分离株，所有碳青霉烯酶均被检测到。开发了一种快速可靠的方法，用于检测和区分德国最常见的 12 种碳青霉烯酶组。该方法允许在进行种属鉴定之前快速测试临床分离株，并且不需要事先进行表型特征分析，是医院感染管理的一种快速而有价值的工具。
