Rapid purification of avian influenza virus for use in enzyme-linked immunosorbent assay.
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% (w/v) 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 enzyme-linked immunosorbent assay. A significant increase in specificity and sensitivity of the enzyme-linked immunosorbent assay 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.
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000302:isolation & purification; D000956:Antigens, Viral / Q000302:isolation & purification*; D002642:Chick Embryo; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006735:Horseradish Peroxidase; D007124:Immunoenzyme Techniques; D009980:Influenza A virus / Q000276:immunology / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis; D011817:Rabbits / Q000276:immunology; D014422:Turkeys / Q000276:immunology
D000888:Antibodies, Anti-Idiotypic; D000956:Antigens, Viral; D006735:Horseradish Peroxidase