title: Successful capture of Toxocara canis larva antigens from human serum samples.
abstract: BACKGROUND
  Toxocara canis is a nematode that parasitizes dogs, while humans are paratenic hosts. When humans are infected the migrating larvae damage the liver, lungs and even the nervous system. Larva migrans diagnosis is based on immunological techniques; however, the commercial immunodiagnostic kits detect anti-T. canis antibodies which may cross-react with other parasites, mainly nematodes with extra-intestinal migration. Moreover, antibodies do not necessarily reflect an active infection; so detection and quantification of circulating antigens may provide appropriate and timely information for treatment, which prevents irreversible damage. Here we report the standardization of a monoclonal antibody based antigen capture ELISA to diagnose human toxocariasis without cross-reaction.
  
  
  METHODS
  We developed anti-T. canis polyclonal antibodies in rabbits and a monoclonal antibody in mouse which did not cross-react with 15 antigens from several parasites. The sandwich ELISA standardization was performed using sera from mice experimentally infected. We tested the method using 29 positive and 58 negative human sera previously typified with a commercial kit, which detects antibodies.
  
  
  RESULTS
  Only 5.0 μg/mL and 10 μg/mL polyclonal antibodies and monoclonal antibody, respectively, were needed in the sandwich ELISA standardization, detecting since 440 pg/mL larva antigens. Nine out of 29 antibody-positive sera were also positive for antigens and no false positive were found. Taking the antibody kit as the reference standard, the sensibility and specificity of the antigen test were 31% and 100%, respectively.
  
  
  CONCLUSIONS
  With these tools we established a detection threshold as low as 440 pg/mL antigen. Monoclonal antibody is specific, and did not cross-react with antigens from other parasites. Detection of circulating antigens helps provide appropriate and timely treatment and prevents irreversible damage.
authors.affiliation: Laboratorio de Parasitología Experimental, Instituto Nacional de Pediatría, Insurgentes Sur No. 3700-C, Colonia Insurgentes Cuicuilco. Delegación Coyoacan, México D.F, 04530, México. bioarca@gmail.com.
authors.forename: Aarón
authors.initials: A
authors.lastname: Rodríguez-Caballero
authors.affiliation: Laboratorio de Parasitología Experimental, Instituto Nacional de Pediatría, Insurgentes Sur No. 3700-C, Colonia Insurgentes Cuicuilco. Delegación Coyoacan, México D.F, 04530, México. marionmgordillo@yahoo.com.
authors.forename: Mario Noé
authors.initials: MN
authors.lastname: Martínez-Gordillo
authors.affiliation: Laboratorio de Anticuerpos Monoclonales, Instituto de Diagnóstico y Referencia Epidemiológicos, Calle Francisco P Miranda No. 177. Col. Unidad Lomas de Plateros, Delegación Álvaro Obregón, México D.F, 01480, México. medinafy@yahoo.com.
authors.forename: Yolanda
authors.initials: Y
authors.lastname: Medina-Flores
authors.affiliation: Laboratorio de Anticuerpos Monoclonales, Instituto de Diagnóstico y Referencia Epidemiológicos, Calle Francisco P Miranda No. 177. Col. Unidad Lomas de Plateros, Delegación Álvaro Obregón, México D.F, 01480, México. medinaeme@yahoo.com.
authors.forename: María Edith
authors.initials: ME
authors.lastname: Medina-Escutia
authors.affiliation: Laboratorio de Pruebas Rápidas, Instituto de Diagnóstico y Referencia Epidemiológicos, Calle Francisco P Miranda No. 177. Col. Unidad Lomas de Plateros, Delegación Álvaro Obregón, México D.F, 01480, México. anmez@hotmail.com.
authors.forename: Antonio
authors.initials: A
authors.lastname: Meza-Lucas
authors.affiliation: Laboratorio de Inmunología Experimental, Instituto Nacional de Pediatría, Insurgentes Sur No. 3700-C, Colonia Insurgentes Cuicuilco. Delegación Coyoacan, México D.F, 04530, México. mariadol@yahoo.com.
authors.forename: Dolores
authors.initials: D
authors.lastname: Correa
authors.affiliation: Laboratorio de Parasitología Experimental, Instituto Nacional de Pediatría, Insurgentes Sur No. 3700-C, Colonia Insurgentes Cuicuilco. Delegación Coyoacan, México D.F, 04530, México. caballerosalazars@gmail.com.
authors.forename: Silvia
authors.initials: S
authors.lastname: Caballero-Salazar
authors.affiliation: Laboratorio de Parasitología Experimental, Instituto Nacional de Pediatría, Insurgentes Sur No. 3700-C, Colonia Insurgentes Cuicuilco. Delegación Coyoacan, México D.F, 04530, México. macotelam@yahoo.com.
authors.forename: Martha
authors.initials: M
authors.lastname: Ponce-Macotela
mesh_terms: D000818:Animals; D000947:Antigens, Helminth / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D007814:Larva / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D017166:Toxocara canis / Q000276:immunology / Q000302:isolation & purification*; D014120:Toxocariasis / Q000097:blood* / Q000175:diagnosis / Q000276:immunology / Q000469:parasitology
chemical_list: D000947:Antigens, Helminth