Serodiagnostic reliability of single-step enriched low-molecular weight proteins of Taenia solium metacestode of American and Asian isolates.
The low-molecular weight proteins (LMWPs) of Taenia solium metacestode (TsM) constituted pertinent serodiagnostic antigens for cysticercosis. We established a novel single-step purification of the LMWPs from TsM cyst fluid (CF). When the CF was precipitated with trichloroacetic acid/acetone mixture at the final concentrations of 5 and 50%, most LMWPs (ranging 7-38kDa) remained in the supernatant fraction. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analyses revealed that the LMWPs consisted mainly with the previously reported subunit proteins of the 120- and 150-kDa antigen complexes. Serum samples from neurocysticercosis (NC) and other helminthic infections, as well as those from healthy controls were tested by both immunoblotting and micro-ELISA. In 135 cases of active stage NC patients, 132 cases (97.7%) showed positive reactions. Serum samples from other helminthic diseases (n=125) and healthy controls (n=100) exhibited no positive reactions except for cystic echinococcosis, of which 12% (3/25 cases) exhibited low levels of cross-reactivity. The LMWPs from different geographical regions (Korea and Mexico) showed diagnostic sensitivity and specificity of 97.7% and 98.7% against active stage NC. Our single-step separation method for the LMWPs provided excellent performance with easy applicability and high reproducibility, which has a great benefit for preparation of potent antigen in endemic areas.
Lee
E-G
EG
Department of Molecular Parasitology, Sungkyunkwan University School of Medicine, Suwon 440-747, Korea.
Bae
Y-A
YA
Kim
S-H
SH
Díaz-Camacho
S P
SP
Nawa
Y
Y
Kong
Y
Y
D000818:Animals; D000947:Antigens, Helminth / Q000276:immunology / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007723:Korea; D008800:Mexico; D008970:Molecular Weight; D020019:Neurocysticercosis / Q000175:diagnosis*; D015203:Reproducibility of Results; D012698:Serologic Tests; D041201:Taenia solium / Q000276:immunology*
D000947:Antigens, Helminth