A Case of Identity Confirmation of Brucella abortus S99 by Phage Typing and PCR Methods

Document Type: Original Articles

Authors

1 Department of Brucellosis, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran

2 Department of Bio bank, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran

3 Department of Brucellosis, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran, Iran

Abstract

Brucellosis is a zoonotic infection that is associated with fever in humans and abortion in animals. The agent of this disease is a facultative intracellular gram-negative coccobacillus called Brucella. There are six classic species, including B. abortus, B. melitensis, B. suis, B. canis, B. neotomae, and B. ovis.  In recent years, four new species have been reported, including Brucella ceti, B. microti, B. pinnipedialis, and B. inopinata. Human disease causes hygienic and economic losses, including inactivity of workforces in the community and high cost of treatment. The disease also causes catastrophic losses in the livestock industry. There is no effective vaccine against human brucellosis. Hence, attempts to prevent human infection with Brucella are focused on preventative measures, including control of infection in livestock, which lead to a reduction in its incidence in humans. The common methods for diagnosis of this disease are serologic methods including Rose Bengal, Wright -2 ME and the ring test. B. abortus strain S99 is used to produce these diagnostic antigens. The production of these antigens requires the presence of a well-characterized seed with full identity. The aim of this work was confirmation of the identity of B. abortus S99 by phage typing, AMOS and multiplex PCR techniques. Therefore, it is essential to carry out the identification of the strains used as seed for the production of the brucellosis diagnostic antigens. In this project, B. abortus strain 99 was supplied by the bacterial collection of the Brucellosis Department of Razi Vaccine and Serum Research Institute. Then, the main aim of the present study was the confirmation of the seed identity by doing the tests through the standard phage typing method, AMOS PCR and multiplex PCR (Brucladder) methods. Results were in support of the identity of the studied strain, and the molecular methods could also be used as the sensitive approaches for validation of antigenic seed.

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Main Subjects


Article Title [French]

Un cas de Confirmation d'Identité de Brucella abortus S99 par Typage du Phage et Méthodes PCR

Abstract [French]

La brucellose est une zoonose à l’origine de fièvre chez l'homme et d’avortement chez l'animal. L'agent de cette maladie est un coccobacille Gram-négatif intracellulaire facultatif appelé Brucella. Il existe six espèces principales, à savoir B. abortus, B. melitensis, B. suis, B. canis, B. neotomae et B. ovis. Ces dernières années, quatre nouvelles espèces ont été signalées, à savoir Brucella ceti, B. microti, B. pinnipedialis et B. inopinata. Chez l’homme, cette maladie est souvent débilitante entraînant des pertes économiques, considérables. La maladie provoque également des pertes catastrophiques dans le secteur de l'élevage. Il n'y a pas de vaccin efficace contre la brucellose humaine. Par conséquent, les tentatives de prévention des infections zoonotiques sont axées sur des mesures préventives dans le but de contrôler l'infection chez le bétail afin de réduire son incidence chez l'homme. Les méthodes habituelles de diagnostic de cette maladie sont les méthodes sérologiques, notamment l’agglutination de Rose Bengal, Wright -2 ME et le ring-test. La souche S99 de B. abortus est utilisée pour produire les antigènes dédiés au diagnostic. La production de ces antigènes nécessite la présence d'une souche bien caractérisée et pleinement identitaire. Le but de ce travail était de confirmer l’identitéde B. abortus S99 par typage sur phage, par les techniques de PCR AMOS et Multiplex. Par conséquent, il est essentiel de procéder à l'identification des souches utilisées pour la production des antigènes utilisés dans le diagnostiques de la brucellose. La souche 99 de B. abortus étudiée dans ce projet provenait de la collection bactérienne du département de brucellose de l’Institut de recherche sur le vaccin et le sérum de Razi. L'objectif principal était ensuite de confirmer l'identité de cette souche en effectuant une batterie de tests parlysotypage, PCR AMOS et PCR multiplex (Brucladder). Les résultats ont confirmé l'identité de la souche étudiée et les méthodes moléculaires présentées dans cet article constituent une approche sensible pour la validation des souches dédiées à la production d’antigénes.

Keywords [French]

  • Test moléculaire
  • Souche antigénique
  • Brucella abortus
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