Development of a highly sensitive IgG-ELISA based on recombinant arginine kinase of Toxocara canis for serodiagnosis of visceral larva migrans in the murine model.
Toxocariasis is a worldwide zoonotic disease caused by the ascarid nematode Toxocara canis. The most common method available for serodiagnosis of toxocariasis is an enzyme-linked immunosorbent assay (ELISA) test using Toxocara excretory-secretory antigen (TES). The present study describes the development of IgG-ELISA based on antiserum prepared against the recombinant arginine kinase of Toxocara canis. Antiserum was prepared against the purified recombinant arginine kinase (AK) using 6-week-old female Japanese white rabbits. Serum samples were collected from experimentally infected BALB/c and C57BL/6 mice at different time periods. The IgG-ELISA was performed using serum samples from mice (infected/uninfected) and TES antigen with antiserum prepared against the recombinant-AK. The optical density (OD450) was measured at 450 nm using a micro-plate ELISA reader. There were significant differences (P<0.01) in the absorbance between infected and control serum samples. Further, we obtained 100% sensitivity for the serum samples from T. canis-infected mice. Therefore, it is suggested that the recombinant-AK based IgG-ELISA could be applied for immunodiagnosis of human toxocariasis. However, it is necessary to evaluate the specificity of this recombinant antigen with similar geohelminth infections.
Wickramasinghe
Susiji
S
Department of Environmental Health Sciences, Kochi Medical School, Oko, Nankoku City, Kochi 783-8505, Japan.
Yatawara
Lalani
L
Nagataki
Mitsuru
M
Takamoto
Misa
M
Watanabe
Yoshiya
Y
Rajapakse
R P V J
RP
Uda
Kouji
K
Suzuki
Tomohiko
T
Agatsuma
Takeshi
T
D000818:Animals; D000909:Antibodies, Helminth / Q000302:isolation & purification; D000947:Antigens, Helminth / Q000097:blood* / Q000235:genetics; D001122:Arginine Kinase / Q000097:blood* / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D007816:Larva Migrans, Visceral / Q000175:diagnosis*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D012698:Serologic Tests; D017166:Toxocara canis / Q000276:immunology*; D014120:Toxocariasis / Q000175:diagnosis
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D001122:Arginine Kinase