Evaluation of Multiplex Real-time Polymerase Chain Reaction in Detecting Common Species of Brucella

Arabestani, Mohammad Reza and Gholami, Ali and Alikhani, Mohammad Yousef and Bahmani, Nasrin and Hashemi, Seyed Hami (2017) Evaluation of Multiplex Real-time Polymerase Chain Reaction in Detecting Common Species of Brucella. Journal of Mazandaran University of Medical Sciences, 27 (147). pp. 97-107.

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Official URL: http://jmums.mazums.ac.ir/article-1-9819-en.html

Abstract

Background and purpose: Brucellosis is a zoonotic disease. Given that serologic test has some specific characteristics such as difficult growth, problems associated with culture, and cross-reactions with otherbacteria, in this study multiplex real-time polymerase chain reaction (PCR) method was used to detect the most common species of Brucella spp. Materials and methods: This experimental study was performed in Hamedan University of Medical Sciences, Hamedan, Iran, 2015, in two bioinformatical and analytical stages. A universal primer was designed to detect Brucella genus and two primers were prepared for B. melitensis and B. abortus, which are the most common species. The target genes for universal Brucella, B. melitensis, and B. abortus primers were ITS, BMEII-0466, and BruAB2-0168, respectively. In the analytical stage, specificity and sensitivity of the primers were evaluated. Results: In this study, the universal primer of Brucella was detected in 83 bp at melt temperature of 82�C. Moreover, B. melitensis and B. abortus primers were specifically identified in 121 bp and 285 bp at melt tempretures of 84�C and 87�C, respectively. The sensitivity for the universal Brucella primer and B. abortus was 50 fg, while it was 100 fg for the B. melitensis primer. Conclusion: The identified primers revealed high levels of sensitivity and specificity. Therefore, they can be considered along with culture in clinical trials.

Item Type: Article
Uncontrolled Keywords: B. abortus, B. melitensis, multiplex, real-time PCR
Depositing User: Unnamed user with email eprints@mazums.ac.ir
Date Deposited: 04 Jan 2018 18:54
Last Modified: 04 Jan 2018 18:54
URI: http://eprints.mazums.ac.ir/id/eprint/1871

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