Serodiagnosis of toxoplasmosis by using a recombinant form of the 54-kilodalton rhoptry antigen expressed in Escherichia coli.
A 330-residue carboxy-terminal antigenic fragment of the Toxoplasma gondii 54-kDa rhoptry protein (ROP2) was expressed in Escherichia coli as a fusion polypeptide containing a 48-amino-acid sequence derived from phage lambda protein Cro and E. coli protein LacI followed by six consecutive histidyl residues. Metal chelate affinity chromatography provided an easy way to isolate the recombinant product in a highly purified form (> 95%). When this material was used to develop an immunoglobulin G (IgG) enzyme-linked immunosorbent assay for diagnosis of toxoplasmosis, the test reached a sensitivity of 89%. The sensitivity of the assay was similar whether the sera contained T. gondii-specific IgM or were devoid of such IgM. It was also found that ROP2 is a conserved antigen since antibodies against the recombinant antigen could be detected in mice experimentally infected with 11 independent T. gondii isolates originating from infected human tissues tested. Thus, the 54-kDa rhoptry antigen could advantageously complement other previously described T. gondii antigens for the development of more-sensitive and more-informative recombinant antigen-based tests for toxoplasmosis diagnosis.
van Gelder
P
P
Innogenetics N.V., Zwijnaarde, Belgium.
Bosman
F
F
de Meuter
F
F
van Heuverswyn
H
H
Hérion
P
P
D000818:Animals; D000953:Antigens, Protozoan / Q000276:immunology / Q000302:isolation & purification; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli; D006801:Humans; D051379:Mice; D010957:Plasmids; D011994:Recombinant Proteins / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D014122:Toxoplasma / Q000276:immunology; D014123:Toxoplasmosis / Q000175:diagnosis* / Q000276:immunology
D000953:Antigens, Protozoan; D011994:Recombinant Proteins