Department of Microbiology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Microbiology, Faculty of Medicine, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
BMC Microbiol. 2019 Jun 10;19(1):122. doi: 10.1186/s12866-019-1510-y.
Acinetobacter baumannii is a cosmopolitan bacterium that is frequently reported from hospitalized patients, especially those patients who admitted in the intensive care unit. Recently, multiplex real-time PCR has been introduced for rapid detection of the resistance genes in clinical isolates of bacteria. The current study aimed to develop and evaluate multiplex real-time PCR to detect common resistance genes among clinical isolates of A. baumannii.
Multiplex real-time PCR based on melting curve analysis showed different T corresponding to the amplified fragment consisted of 83.5 °C, 93.3 °C and 89.3 °C for blaIMP, blaVIM and blaNDM, respectively. Results of multiplex real-time PCR showed that the prevalence of blaIMP, blaVIM and blaNDM among the clinical isolates of A. baumannii were 5/128(3.9%), 9/128(7.03%) and 0/128(0%), respectively. Multiplex real-time PCR was able to simultaneously identify the resistance genes, while showed 100% concordance with the results of conventional PCR.
The current study showed that blaVIM, was the most prevalent MBL gene among the clinical isolates of A. baumannii while no amplification of blaNDM was seen. Multiplex real-time PCR can be sensitive and reliable technique for rapid detection of resistance genes in clinical isolates.
鲍曼不动杆菌是一种广泛存在的细菌，经常在住院患者中报告，尤其是那些住进重症监护病房的患者。最近，多重实时 PCR 已被引入用于快速检测临床分离菌中的耐药基因。本研究旨在开发和评估多重实时 PCR 以检测临床分离的鲍曼不动杆菌中的常见耐药基因。
基于熔解曲线分析的多重实时 PCR 显示，与扩增片段对应的 T 值不同，分别为 blaIMP、blaVIM 和 blaNDM 的 83.5°C、93.3°C 和 89.3°C。多重实时 PCR 的结果显示，临床分离的鲍曼不动杆菌中 blaIMP、blaVIM 和 blaNDM 的流行率分别为 5/128(3.9%)、9/128(7.03%)和 0/128(0%)。多重实时 PCR 能够同时鉴定耐药基因，与常规 PCR 的结果具有 100%的一致性。
本研究表明，blaVIM 是临床分离的鲍曼不动杆菌中最常见的 MBL 基因，而未见 blaNDM 的扩增。多重实时 PCR 是一种敏感可靠的技术，可用于快速检测临床分离菌中的耐药基因。
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