Rapid purification of avian influenza virus for use in enzyme-linked immunosorbent assay
Abraham, A.; Sivanandan, V.; Newman, J.A.; Maheswaran, S.K.
American Journal of Veterinary Research 45(5): 959-962
1984
ISSN/ISBN: 0002-9645 PMID: 6375486 Document Number: 239404
A rapid and easy purification method was developed to obtain avian influenza antigen for use in immunochemical assays. This was achieved by rapid concentration of virus from infective allantoic fluid, using 8% (wt/vol) polyethylene glycol 8000 and later by purification on gel-permeation chromatography through controlled-pore glass beads. Rabbit anti-turkey globulins were made specific for turkey globulins using affinity chromatography, conjugated to horseradish peroxidase and used in ELISA. A significant increase in specificity and sensitivity of the ELISA was observed when purified antigen was used in place of a crude antigen preparation. This purified antigen eliminated the false-positives obtained as a result of the turkeys being previously vaccinated with egg-grown virus vaccines (Newcastle disease virus). The details of the technique and the importance of antigen preparation are discussed [The purified antigen has application in serodiagnosis of avian influenza.].