Serodiagnosis of toxoplasma infection using a purified parasite protein (P30).
The major surface protein (P30) of Toxoplasma gondii has been purified by immunoabsorption with anti-P30 monoclonal antibodies linked to a glutardialdehyde activated affinity absorbant. SDS-PAGE analysis of the eluted material followed by silver staining showed only a single band of 30,000 mol wt. Western blotting using antibodies from a rabbit immunized with purified P30 against the total Toxoplasma extract separated by SDS-PAGE again revealed an unique antigen of 30,000 daltons. The presence of repeated epitopes within P30 was confirmed by a two-site/one-antibody radiometric assay with the purified protein. Sandwich ELISA procedures with purified P30 clearly demonstrated that all 37 tested patients with acute toxoplasmosis presented significantly high levels of IgM anti-P30 antibodies. In addition, all 40 tested patients with chronic toxoplasma infection also showed high IgG anti-P30 antibody levels. These findings represent an essential step for the development of new reagents for the diagnosis of toxoplasmosis.
F
F
Santoro
D
D
Afchain
R
R
Pierce
J Y
JY
Cesbron
G
G
Ovlaque
A
A
Capron
D000953:Antigens, Protozoan / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D006801:Humans; D007075:Immunoglobulin M / Q000096:biosynthesis; D008565:Membrane Proteins / Q000302:isolation & purification; D015800:Protozoan Proteins; D014122:Toxoplasma / Q000276:immunology; D014123:Toxoplasmosis / Q000175:diagnosis*
D000953:Antigens, Protozoan; D000939:Epitopes; D007075:Immunoglobulin M; D008565:Membrane Proteins; D015800:Protozoan Proteins; C047168:SAG1 antigen, Toxoplasma