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

Document Type : Original Articles

Authors

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

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

Main Subjects

Article Title [French]

Développement d'une Nouvelle Réaction en Chaîne de Polymérase Simple Modifiée et d'une Réaction en Chaîne de Polymérase en Temps Réel pour l'identification des Souches Iraniennes de Brucella abortus

Abstract [French]

La brucellose est principalement une maladie zoonotique dans le monde entier causée par l'espèce Brucella. Le genre Brucella contient des espèces hautement infectieuses classées comme agents de menace biologique. À cet égard, l'identification de Brucella peut être un processus long et exigeant en main-d'œuvre, présentant un risque réel d'infection acquise en laboratoire pour le personnel de laboratoire. Cette étude visait à présenter un nouveau test de réaction en chaîne par polymérase (PCR) classique et en temps réel pour l'identification des souches de Brucella abortus. À cet égard, deux amorces (bru ab2) ont été conçues sur la base des locus uniques codant pour une protéine contenant une répétition de brin bêta associée à un autotransporteur (ID: YP00113760). Un total de 56 souches de Brucella (par exemple, des isolats de référence, vaccinal et de terrain) et Yersinia enterocolitica, en tant qu'isolât non-Brucella, ont été évalués dans des systèmes de PCR conventionnels et en temps réel. Les résultats de l’étude ont indiqué que l’ADN génomique des souches de B. abortus à 0,4 ng et 400 FG peut être détecté par PCR conventionnelle et en temps réel, respectivement. Les amorces, bru ab2, convenaient aux deux méthodes de PCR. Les deux méthodes étaient spécifiques à la détection de toutes les souches de la bactérie; cependant, le test de PCR en temps réel était 1000 fois plus sensible que le procédé PCR classique. Par conséquent, ce nouveau système de détection pourrait constituer une méthode modifiée sélective et appropriée pour l'identification précise de toutes les souches de B. abortus.

Keywords [French]

  • Brucella abortus
  • identification
  • PCR
  • PCR en Temps Réel

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Archives of Razi Institute
Volume 74, Issue 3
September 2019
Pages 235-241

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