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靶向基因的传统PCR和实时PCR作为快速检测耐碳青霉烯临床菌株的可靠方法

Conventional and Real-Time PCR Targeting Genes as Reliable Methods for a Rapid Detection of Carbapenem-Resistant Clinical Strains.

作者信息

Depka Dagmara, Mikucka Agnieszka, Bogiel Tomasz, Rzepka Mateusz, Zawadka Patryk, Gospodarek-Komkowska Eugenia

机构信息

Microbiology Department, Ludwik Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toruń, 85-094 Bydgoszcz, Poland.

出版信息

Antibiotics (Basel). 2022 Mar 28;11(4):455. doi: 10.3390/antibiotics11040455.

Abstract

Multidrug-resistant , particularly those producing carbapenemases, are spread worldwide. A reliable detection of carbapenemases is essential to choose the appropriate antimicrobial therapy and, consequently, prevent the dissemination of carbapenem-resistant strains. The aim of this study is to examine the molecular basis of the carbapenem resistance mechanism and estimation of conventional PCR and real-time PCR usefulness for the detection of oxacillinases when compared to phenotypic carbapenemases detection. The following methods were evaluated: the CarbAcineto NP test, Carbapenem Inactivation Method, CPO panels of semiautomated antimicrobial susceptibility testing method on the BD Phoenix™ M50 system, conventional Polymerase Chain Reaction and real-time PCR. The eazyplex SuperBug complete A assay was used as the reference method. Among the tested strains, 39 (67.2%) carried the gene, while the gene was noted amongst 19 (32.8%) isolates. The diagnostic sensitivities of the studied assays were as follows: CarbAcineto NP-65.5%; CIM-100%; CPO-100%; conventional PCR-100%; real-time PCR-100%.

摘要

多重耐药菌,尤其是那些产生碳青霉烯酶的菌株,在全球范围内传播。可靠地检测碳青霉烯酶对于选择合适的抗菌治疗方法并进而防止耐碳青霉烯菌株的传播至关重要。本研究的目的是研究碳青霉烯耐药机制的分子基础,并评估与表型碳青霉烯酶检测相比,传统PCR和实时PCR检测奥沙西林酶的有效性。评估了以下方法:CarbAcineto NP试验、碳青霉烯灭活法、BD Phoenix™ M50系统上的半自动抗菌药物敏感性检测方法的CPO板、传统聚合酶链反应和实时PCR。eazyplex SuperBug complete A检测用作参考方法。在所测试的菌株中,39株(67.2%)携带该基因,而在19株(32.8%)分离株中发现了该基因。所研究检测方法的诊断敏感性如下:CarbAcineto NP为65.5%;CIM为100%;CPO为100%;传统PCR为100%;实时PCR为100%。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1a74/9032895/25cd4e952229/antibiotics-11-00455-g001.jpg

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