Igm recognition of recombinant Toxoplasma gondii antigens by sera of acutely or latently infected humans.
Clinical non-relevant (CNR) IgM specific for Toxoplasma gondii is responsible for false-positive results in commercially available IgM assays. Using IgM immunoblotting, it is possible to distinguish between IgM in sera of acutely infected (AI) patients and CNR IgM. Especially the combination of staining of a 55 and 30 kD antigen in T.gondii lysate proved useful in this respect. The 55 kD antigen was identified as Rop1, while the 30 kD antigen was confirmed to be Sag1. However, the use of recombinant antigens instead of lysates for diagnostic assays would improve reproducibility. IgM recognized recombinant Rop1, but most CNR sera also had low anti-Rop1 titers. Although purified native Sag1 separated AI and CNR sera very well on immunoblot, IgM did not recognize recombinant Sag1 at all. Clearly, it is difficult to produce a recombinant Sag1 that can be recognized by IgM. Recombinant Rop1 might be suitable as one of the recombinant antigens in an IgM immunoblot assay, but has to be combined with at least one other immunogenic antigen.
Meek
Bob
B
Department of Molecular Immunology, The Netherlands Ophthalmic Research Institute, Amsterdam, The Netherlands.
Diepersloot
Robert Jan
RJ
van Gool
Tom
T
Speijer
Dave
D
Peek
Ron
R
D000208:Acute Disease; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology; D000953:Antigens, Protozoan / Q000032:analysis* / Q000276:immunology; D001483:Base Sequence; D005260:Female; D006801:Humans; D015151:Immunoblotting; D007075:Immunoglobulin M / Q000032:analysis / Q000276:immunology*; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D014122:Toxoplasma / Q000276:immunology*; D014123:Toxoplasmosis / Q000097:blood / Q000175:diagnosis*
D000888:Antibodies, Anti-Idiotypic; D000953:Antigens, Protozoan; D007075:Immunoglobulin M; D011994:Recombinant Proteins