Rapid immunofiltration assay of Newcastle disease virus using a silicon sensor.
A rapid nonradioactive sandwich immunoassay which utilizes biotin-streptavidin mediated filtration capture of immune complexes in conjunction with a silicon sensor was developed for the detection of virus. Using purified Newcastle disease virus as a model, the lower limits of detection (LOD) were determined for a number of immunoassay configurations employing both monoclonal and polyclonal antibodies. The LODs ranged from 1.3 ng/ml (sample volume of 100 microliter) for an incubation of 60 min to 400 ng/ml for a 1 min incubation. The sandwich immune complexes were formed from one-step incubation of antibody and antigen. No 'hook' effects were observed over a wide range of analyte concentrations. The assays were easy to perform and required a total time equal to the incubation period plus about 5 min. The assay format is suitable for virus, bacteria and protein antigens. New assays can be developed and optimized readily, often within 1 day.
Lee
W E
WE
Defense Research Establishment Suffield, Medicine Hat, Alberta, Canada.
Thompson
H G
HG
Hall
J G
JG
Fulton
R E
RE
Wong
J P
JP
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D001426:Bacterial Proteins; D015374:Biosensing Techniques; D001710:Biotin; D005069:Evaluation Studies as Topic; D005374:Filtration / Q000379:methods; D019793:Fluorescein; D005452:Fluoresceins; D007118:Immunoassay / Q000379:methods*; D009522:Newcastle disease virus / Q000276:immunology / Q000302:isolation & purification*; D012825:Silicon; D019809:Streptavidin; D014510:Urease
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D001426:Bacterial Proteins; D005452:Fluoresceins; D001710:Biotin; D019809:Streptavidin; D014510:Urease; D019793:Fluorescein; D012825:Silicon