http://cdjournal.muk.ac.ir/index.php/cdj The Chronic Diseases Journal, a quarterly peer-reviewed scientific journal published by Kurdistan University of Medical Sciences since 2013, is certified by the Medical Journals Commission of the Iranian Ministry of Health and Medical Education as a SCIENTIFIC-RESEARCH journal (No. 1392/04/20-15/92/4508) since 2019. It publishes approximately 32-40 research articles and 2-3 review articles annually, with a submission-to-first decision time of 2 days, a submission-to-final decision time of 3 weeks, and an acceptance-to-publication time of 2-10 weeks. Vesnu Publications en-US Chronic Diseases Journal 2588-7297 <p>This work is licensed under a <a href="https://creativecommons.org/licenses/by-nc/4.0/" target="_blank" rel="noopener">Creative Commons Attribution-NonCommercial 4.0</a> Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.</p> http://cdjournal.muk.ac.ir/index.php/cdj/article/view/966 BACKGROUND: Diagnosis of brucellosis requires a rapid and accurate method such as the polymerase chain reaction (PCR). The purpose of this study was simultaneous detection of Brucella abortus (B. abortus) and Brucella melitensis (B. melitensis) in serum samples using the duplex PCR technique and then comparing the results using the Rose Bengal test (RBT). METHODS: In this comparative-descriptive study, 100 serum samples were collected from a veterinary station located in Shahriar City, Iran. Moreover, the monoplex-PCR of B. abortus and B. melitensis and duplex-PCR for both agents was optimized. The limit of detection (LOD) and specificity test were also checked. Besides, deoxyribonucleic acids (DNAs) were extracted from the serum samples by the DNA extraction solution (DNG-plus) technique. The PCR product was cloned in pTZ57R plasmid by T/A cloning. RESULTS: B. abortus (494bp) and B. melitensis (733bp) amplicons were observed in 1.5% gel electrophoresis. The LOD of the monoplex-PCR test for both of the agents was 100 genomes per reaction. Additionally, 40 out of 100 samples were positive for RBT, out of them, 35 samples were positive with duplex-PCR, 31 samples were positive for B. abortus, and 4 for B. melitensis; moreover, 20 samples were positive with duplex PCR from 60 negative RBT. From this number, 17 samples of B. abortus and 3 samples of B.melitensis were detected. CONCLUSION: The number of positive samples by duplex-PCR was more than the RBT; therefore, we can assert duplex-PCR for confirming the RBT results. Shabnam Bahrami Pegah Shakib Mohammad Reza Zolfaghari Mohammad Hassan Shahhosseini Sanaz Ahmadi Copyright (c) 2024 Chronic Diseases Journal https://creativecommons.org/licenses/by-nc/4.0 2025-02-04 2025-02-04 26 32 10.22122/cdj.v13i1.966