International Journal of Microbiology and Immunology Research Vol. 3(1), pp. 009-010, February 2015 ISSN 2327-7769 ©2015 Academe Research Journals

Short Communication

Identification of infectious bursal disease virus (IBDV) through agar gel immunodiffusion (AGID) test and its purification by ultracentrifugation

Tariq Mehmood Butt, Rehan Rafique*, Muhammad Najiullah khan, Talha Farooq, Umber Rauf, Yasmeen Abbas, Muhammad Asad Raza and Asaad Nayyer

Veterinary Research Institute, Zarrar Shaheed Road, Lahore Cantt. Punjab, Zip Code/Postal Code 54810, Pakistan.

*Corresponding author. E-mail: dr.rehan702@gmail.com. Tel: +92-333-6607756.

Accepted 6 January, 2015

Abstract

Infectious bursal disease (IBD) is a highly important immunosuppressive disease of young chickens caused by infectious bursal disease virus (IBDV). Infected bursae were collected from filed outbreaks of IBD at Lahore, Pakistan. A 10% W/V suspension was made in the phosphate buffer saline and centrifuged at 5000 rpm for 20 min. The presence of IBDV in the supernatants of suspect homogenates was checked through agar gel immuno-diffusion test. The collected material was given serial passage in 10 days old embryonating eggs at an inoculation rate of 0.2 ml/egg through chroio-allontoic membrane (CAM) route. The material collected at different passages was centrifuged twice at 20000 rpm for 45 min to remove tissue debris. The clear supernatant was subjected to ultracentrifugation at 49500 × g for 6 h at 4°C. The purified IBD virus was obtained in pellet form and these pellets were disrupted in TNE buffer to be stored at -70°C till further use. The study suggested that the bursa of fabricius is the best tissue source for the isolation of virus.

Key words: Young chicken, infected bursae, isolation of IBD virus, agar gel immuno-diffusion (AGID), ultracentrifugation.