Dot-blot ELISA for the detection of IgM RF and IgA RF.
Dot-blot ELISA was developed for the detection of IgM RF and IgA RF. Normal rabbit IgG (NRIgG), concentration 100 micrograms/ml, was used as the antigen for dotting on the 0.45 microns pore size nitrocellulose membrane. Serum, conjugate and substrate incubation conditions were at room temperature for 1 hour, 1 hour and 3 minutes, respectively. The membrane with NRIgG dot could be sotred for 6 weeks before use in the assay. Positive results of IgM RF, at the serum dilution 1:800, were found in 31/51 patients with either classical or definite rheumatoid arthritis and 3/68 normal healthy individuals. Positive IgA RF, at the serum dilution 1:100, was found in 27/51 of the former and none of the latter. Significant concordance with high agreement index was found between the results of the dot-blot ELISA developed and those obtained from ELISA performed in microtitre plate (Kappa greater than or equal to 0.78 for IgM RF and 0.83 for IgA RF, p less than 0.001).
Banchuin
N
N
Department of Microbiology, Faculty of Medicine, Siriraj Hospital, Bangkok, Thailand.
Janyapoon
K
K
Parivisutt
L
L
Sarntivijai
S
S
D000293:Adolescent; D000328:Adult; D000368:Aged; D001172:Arthritis, Rheumatoid / Q000276:immunology; D002648:Child; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D015151:Immunoblotting; D007070:Immunoglobulin A / Q000097:blood*; D007075:Immunoglobulin M / Q000097:blood*; D008875:Middle Aged; D012217:Rheumatoid Factor / Q000097:blood*
D007070:Immunoglobulin A; D007075:Immunoglobulin M; D012217:Rheumatoid Factor