Production of monoclonal antibody against Toxocara cati second-stage larvae and its application for the detection of circulating antigens.

Toxocariasis is an important zoonotic disease caused by the second-stage larvae of Toxocara species. Human toxocariasis is usually diagnosed by serological methods. We prepared monoclonal antibody specific for Toxocara cati (T. cati) and performed the following experiments using this antibody to increase the sensitivity of our capture ELISA for antigen detection. The isotype of monoclonal antibody was IgG3. Using this monoclonal antibody, a capture ELISA was developed. The produced monoclonal antibody reacted well with excretory-secretory (ES) antigen of T. cati with protein nature but not with other T. cati related antigens. A monoclonal antibody was raised against ES antigen of T. cati larval stage. No cross-reaction was found with antigens of Ascaris suum, Dirofilaria immitis, Trichostrongylus orientalis, encysted larvae of Trichinella spiralis, and metacercariae of Centrocestus armatus. The system had enough sensitivity to detect 5 ng/mL of circulating antigens in the serum. The results indicate that the new antigen detection system provides a useful method for the diagnosis of visceral and ocular larva migrans caused by T. cati and fulfils the requirements of a serological method for diagnosis of toxocariasis.

[1]  W. Roldán,et al.  Evaluation of an enzyme-linked immunoelectrotransfer blot test for the confirmatory serodiagnosis of human toxocariasis. , 2009, Memorias do Instituto Oswaldo Cruz.

[2]  A. Oryan,et al.  Histopathologic changes and larval recovery of Toxocara cati in experimentally infected chickens , 2007, Parasitology Research.

[3]  M. Zibaei,et al.  A Method for Accelerating the Maturation of Toxocara cati Eggs , 2007 .

[4]  R. Maizels,et al.  Comparison of IgG-ELISA and IgG4-ELISA for Toxocara serodiagnosis. , 2005, Acta tropica.

[5]  R. Fabre,et al.  Application of the Western blotting procedure for the immunodiagnosis of human toxocariasis , 2004, Parasitology Research.

[6]  D. Despommier,et al.  Toxocariasis: Clinical Aspects, Epidemiology, Medical Ecology, and Molecular Aspects , 2003, Clinical Microbiology Reviews.

[7]  M. Tsuji,et al.  Sandwich ELISA detection of excretory-secretory antigens of Toxocara canis larvae using a specific monoclonal antibody. , 2002, The Southeast Asian journal of tropical medicine and public health.

[8]  G. X. Wang,et al.  Detection of circulating antigens and antibodies in Toxocara canis infection among children in Chengdu, China. , 1999, The Journal of parasitology.

[9]  S. Fenoy,et al.  Cross-reactions of sera from Toxascaris leonina and Ascaris suum infected mice with Toxocara canis, Toxascaris leonina and Ascaris suum antigens. , 1995, International journal for parasitology.

[10]  S. Fenoy,et al.  Evaluation of chemotherapy in experimental toxocarosis by determination of specific immune complexes , 1990, Journal of Helminthology.

[11]  P. Schantz Toxocara larva migrans now. , 1989, The American journal of tropical medicine and hygiene.

[12]  T. Burkot,et al.  Detection of circulating parasite antigen and specific antibody in Toxocara canis infections. , 1988, Clinical and experimental immunology.

[13]  R. Maizels,et al.  Species‐specific and common epitopes on the secreted and surface antigens of Toxocara cati and Toxocara canis infective larvae , 1987, Parasite immunology.

[14]  R. Maizels,et al.  Shared carbohydrate epitopes on distinct surface and secreted antigens of the parasitic nematode Toxocara canis. , 1987, Journal of immunology.

[15]  D. Bowman,et al.  Circulating excretory-secretory antigen levels and specific antibody responses in mice infected with Toxocara canis. , 1987, The American journal of tropical medicine and hygiene.

[16]  A. W. Woodruff,et al.  Toxocariasis: serological diagnosis by enzyme immunoassay. , 1979, Journal of clinical pathology.

[17]  D. Savigny In vitro maintenance of Toxocara canis larvae and a simple method for the production of Toxocara ES antigen for use in serodiagnostic tests for visceral larva migrans. , 1975, The Journal of parasitology.