Isolation and purification of Echinococcus granulosus antigen B from hydatid cyst fluid using three different methods

Document Type : Original Articles

Authors

1 Department of Pathobiology, Faculty of Veterinary, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Proteomics and Biochemistry Department, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.

3 Department of Microbiology, School of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.

4 Department of Pathobiology, Faculty of Veterinary, Islamic Azad University, Garmsar, Iran

Abstract

Hydatid cyst, the larval stage of cestodes Echinococcus spp., is recognized as a zoonotic infection in the world. The World Health Organization (WHO) has recently classified echinococcosis in a group of neglected tropical diseases.The prevalence of Echinococcus granulosus infection is high in Iran due to the presence of various intermediate hosts in this country. Considering the rising trend of this zoonotic parasitic disease based on national epidemiological studies, diagnosis is of great significance. WHO has suggested the use of specific antigens, especially antigen B (AgB) for serological diagnostic tests. In general, AgB is a polymeric lipoprotein, which disintegrates into 8.12, 16, and 20.24 kDa subunits. In the present study, we applied three different methods for AgB isolation from hydatid cyst fluid (HCF) and compared their efficacy in AgB isolation. Finally, the protein concentration of this antigen was measured by Bradford assay and confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The results showed that the application of polyethylene glycol (PEG 4000) as a thickener agent beside purification of HCF in dialysis bag and filtering and also dialysis against acetate buffer leading to the best quantity in purified antigen B.

Keywords

Main Subjects

Article Title [French]

Isolation et purification de l’antigène B de l’Echinococcus granulosus à partir du fluide d’un kyste hydatique par trois différentes méthodes

Abstract [French]

Le kyste hydatique est une zoonose provoquée par la larve des cestodes du genre Echinococcus dans de nombreuses régions du monde. L’Organisation Mondiale de la Santé (OMS) a récemment classifié l’échinicoccose dans le groupe des maladies tropicales jusqu’alors négligées. En Iran, la prévalence des infections à l’Echinococcus granulosus est importante en raison de l’existence d’hôtes intermédiaires variées à travers le pays. Etant donnée l’augmentation de cette zoonose parasitique soulignée par les études épidémiologiques nationales, son diagnostique est d’une grande importance. L’OMS propose l’utilisation d’antigènes spécifiques, particulièrement l’antigène B (AgB), dans l’élaboration de tests sérologiques. L’AgB est une lipoprotéine polymérique constituée de plusieurs sous-unités de 8.12, 16 et 20.24 kDa. Dans cette étude, les rendements de trois différentes méthodes dans l’isolation de l’AgB à partir du fluide du Kyste hydatique (HCF) ont été comparés. Pour cela, la concentration de l’antigène a été mesurée par la méthode de Bradford et son degré de pureté vérifié par une électrophorèse en gel de polyacrylamide contenant du laurylsulfate de sodium (SDS-PAGE). Nos résultats montrent que l’application du polyéthylène glycol (PEG 4000) comme agent épaississant, complémentée d’une filtration et d’un dialyse contre du tampon acétate (pH 8), contribue à un meilleur rendement de purification de l’HCF.

Keywords [French]

  • Antigène B
  • Echinococcus granulosus
  • Kyste hydatique
  • Isolation

References

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Archives of Razi Institute
Volume 71, Issue 2
May and June 2016
Pages 103-108

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