Quantitative antibody ELISA for leprosy.
Quantitative enzyme-linked immunosorbent assays (ELISAs) were established to measure IgM and IgG antibody levels to soluble Mycobacterium leprae sonicate (CD60) and to the synthetic disaccharide antigen based on the phenolic glycolipid-I antigen of M. leprae coupled to bovine serum albumin in 46 leprosy patients. Separate reference pools for IgM and IgG antibody were established. The reciprocal of the antibody titer was expressed as the number of arbitrary units in the reference pools which was subsequently used as the calibrator for assessment of units in individual test sera. The dose-response relationship for both IgM and IgG was highly specific and reproducible for both isotypes, as indicated by the intra- and inter-assay coefficients of variation. The distribution of antibody levels are in general agreement with the results from previous studies against different M. leprae antigens. The lepromatous group showed 10- to 100-fold higher IgM antibodies to both the soluble sonicate antigen and the disaccharide as compared to the control group. Very low to undetectable levels of IgM antibodies were observed in the tuberculoid group of leprosy patients. IgG antibodies, on the other hand, were not only present but showed considerable overlap with the lepromatous patient group. Optimized ELISAs, such as the one described in this study, would allow one to address issues such as antibody changes with treatment, antigen clearance, and correlation with other immune parameters associated with disease pathogenesis and protection.
Hasan
R
R
Department of Microbiology, Aga Khan University, Karachi Pakistan.
Dockrell
H M
HM
Chiang
T
T
Hussain
R
R
D000907:Antibodies, Bacterial / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis*; D007075:Immunoglobulin M / Q000032:analysis*; D009166:Mycobacterium leprae / Q000276:immunology*; D011786:Quality Control
D000907:Antibodies, Bacterial; D007074:Immunoglobulin G; D007075:Immunoglobulin M