Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 12 2 2020 04 01 107 112 Reza Ranjbar Molecular Biology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran Shahin Zayeri Molecular Biology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran Amir Mirzaie Molecular Biology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran 2019 01 26 2020 02 12 Background and Objectives: Acinetobacter baumannii has been known as a major pathogen causing nosocomial infections. The aim of this study was to develop multiplex PCR for rapid and simultaneous detection of metallo-β-lactamase (MBL) genes in clinical isolates of A. baumannii. Materials and Methods: In this study, we used three sets of primers to amplify the MBL genes including blaOXA-48, blaOXA-23 and blaNDM. The multiplex PCR assay was optimized for rapid and simultaneous detection of MBL genes in A. baumannii strains recovered from clinical samples. Results: A. baumannii strains recovered from clinical samples were subjected to the study. The multiplex PCR produced 3 bands of 501 bp for blaOXA-23, 744 bp for blaOXA-48 and 623 bp for blaNDM genes. In addition to, no any cross-reactivity was observed in multiplex PCR. Conclusion: Based on obtained data, the multiplex PCR had a good specificity without any cross reactivity and it appears that the multiplex PCR is reliable assay for simultaneous detection of MBL genes in A. baumannii strains. https://ijm.tums.ac.ir/index.php/ijm/article/view/2021 https://ijm.tums.ac.ir/index.php/ijm/article/download/2021/1229