Development and field validation of an avidin-biotin enzyme-linked immunosorbent assay kit for bovine brucellosis.
The avidin-biotin enzyme-linked immunosorbent assay (A-B ELISA), for use in surveillance for bovine brucellosis in India was developed and calibrated using the indirect brucellosis ELISA kit of the International Atomic Energy Agency (IAEA) as a reference. The reagents used in the A-B ELISA were as follows: the smooth lipopolysaccharide of Brucella abortus strain 99 (antigen); biotinylated anti-bovine immunoglobulin G (detection antibody); avidin-horseradish peroxidase (conjugate); and O-phenylenediamine dihydrochloride (chromogen). The test results were interpreted using the IAEA software EDI version 2.1.1, which was modified for use in the A-B ELISA. The cut-off percentage positivity value was established using 500 brucellosis-positive and 500 brucellosis-negative serum samples, confirmed with reference to the sample data using the indirect ELISA kit. The overall specificity of A-B ELISA was 98.8% and overall sensitivity was 98.2%. Field validation of the A-B ELISA kit was undertaken in six laboratories in India. Screening of 7,040 cattle and 678 buffalo serum samples from 12 states revealed serological evidence of brucellosis in 8.7% of cattle and 10.2% of buffalo. This kit proved to be robust and performed with a similar sensitivity and specificity to the indirect ELISA. The kit can be supplied at a lower cost than current commercial ELISA kits.
Renukaradhya
G J
GJ
Food and Agriculture Organization/International Atomic Energy Agency (IAEA), Joint Division, IAEA, Wagramerstrasse 5, P.O. Box 100, A-1400 Vienna, Austria.
Isloor
S
S
Crowther
J R
JR
Robinson
M
M
Rajasekhar
M
M
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D001360:Avidin; D001710:Biotin; D002003:Brucella abortus / Q000276:immunology*; D002007:Brucellosis, Bovine / Q000175:diagnosis* / Q000453:epidemiology; D002020:Buffaloes; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D007106:Immune Sera / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology
D000907:Antibodies, Bacterial; D007106:Immune Sera; D008070:Lipopolysaccharides; C082161:lipopolysaccharide A; D001360:Avidin; D001710:Biotin