An ELISA for detection of infectious bursal disease virus and differentiation of very virulent strains based on single chain recombinant chicken antibodies.
Two chicken single-chain variable antibody fragments (scFv) designated scFv154 and scFv88, previously shown to react with either all or very virulent (vv) infectious bursal disease virus (IBDV) strains, respectively, were evaluated for use in an enzyme-linked immunosorbent assay (ELISA) for differentiation of vvIBDV. Specificity and sensitivity of the vvIBDV ELISA was assessed when scFv154 and scFv88 were expressed as soluble antibodies (sAb), phage antibodies (pAb) or hyper-phage antibodies (hpAb). The highest test sensitivity and specificity was obtained using hpAb154 to detect all IBDV and pAb88 to differentiate vvIBDV strains. Such an ELISA was eight to 16 times more sensitive for IBDV antigen detection than the mouse monoclonal antibody ELISA. Using field samples, the scFv ELISA was able to differentiate between flocks infected with vvIBDV and those infected with classical or variant IBDV. In one instance IBDV was detected in a flock found to be negative by the monoclonal antibody ELISA. The results showed that scFv can be utilized as highly specific and sensitive ELISA reagents for the detection and discrimination of avian pathogens.
CSIRO Livestock Industries, Australian Animal Health Laboratory, Private Bag 24, 5 Portarlington Road, Geelong, 3220, Australia.
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S
Sapats
G
G
Gould
L
L
Trinidad
L H
LH
Parede
C
C
David
J
J
Ignjatovic
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology; D018175:Birnaviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D007243:Infectious bursal disease virus / Q000145:classification / Q000276:immunology / Q000302:isolation & purification* / Q000472:pathogenicity*; D011201:Poultry Diseases / Q000175:diagnosis* / Q000821:virology; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D014774:Virulence
D000914:Antibodies, Viral; D000956:Antigens, Viral; D011994:Recombinant Proteins