Development of New Modified Simple Polymerase Chain Reaction and Real-time Polymerase Chain Reaction for the Identification of Iranian Brucella abortus Strains

AUTHORS

S. Alamian 1 , * , T. Zahraei Salehi 2 , K. Aghaiypour Kolyani 3 , M. Esmaelizad 3 , A. Etemadi 1

1 Department of Brucella, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

2 Department of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

3 Department of Biotechnology, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

How to Cite: Alamian S, Zahraei Salehi T, Aghaiypour Kolyani K, Esmaelizad M, Etemadi A. Development of New Modified Simple Polymerase Chain Reaction and Real-time Polymerase Chain Reaction for the Identification of Iranian Brucella abortus Strains, Arch Razi Inst. 2019 ; 74(3):e98537. doi: 10.22092/ari.2018.122128.1218.

ARTICLE INFORMATION

Archives of Razi Institute: 74 (3); 235-241
Published Online: October 01, 2019
Article Type: Journal Article
Received: August 29, 2018
Accepted: September 30, 2018
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Abstract

Brucellosis is primarily a worldwide zoonotic disease caused by Brucella species. The genus Brucella contains highly infectious species that are classified as biological threat agents. In this regard, the identification of Brucella can be a time-consuming and labor-intensive process posing a real risk of laboratory-acquired infection to the laboratory staff. This study aimed to present a novel conventional and real-time polymerase chain reaction (PCR) assay for the identification of Brucella abortus strains. Regarding this, two primers (bru ab2) were designed based on the unique loci encoding autotransporter-associated beta strand repeat-containing protein (ID:YP00113760). A total of 56 Brucella strains (e.g., reference, vaccinal, and field isolates) and Yersinia enterocolitica, as a non-Brucella isolate, were evaluated in conventional and real-time PCR systems. The results of the study indicated that 0.4 ng and 400 FG of genomic DNA of B. abortus strains can be detected by conventional and real-time PCR, respectively. The primers, bru ab2, were suitable for both PCR methods. Both methods were specific for the detection of all strains of the bacterium; however, real-time PCR assay was 1000-fold more sensitive than the conventional PCR method. Therefore, this new detection system could be a suitable selective modified method for the accurate identification of all B. abortus strains.

Keywords

Brucella abortus Identification PCR Real-time PCR

© 2019, Archives of Razi Institute. Razi Vaccine and Serum Research Institute.

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