Browse > Article

Characterization of a Toxocara canis species-specific excretory-secretory antigen(TcES-57) and development of a double sandwich ELISA for diagnosis of visceral larva migrans  

Iddawela, R.D. (Department of Parasitology, Faculty of Medicine, University of Peradeniya)
Rajapakse, R.P.V.J. (Department of Veterinary Pathobiology, Faculty of Veterinary Medicine and Animal Science, University of Peradeniya)
Perera, N.A.N.D. (Department of Veterinary Pathobiology, Faculty of Veterinary Medicine and Animal Science, University of Peradeniya)
Agatsuma, Takeshi (Department of Environmental Health Science, Kochi Medical School)
Publication Information
Parasites, Hosts and Diseases / v.45, no.1, 2007 , pp. 19-26 More about this Journal
Abstract
This study describes the isolation of a Toxocara canis species-specific excretory-secretory(ES) antigen and the development of an enzyme-linked immunosorbent assay(ELISA) based on this antigen. Analysis of the ES antigens of T. canis, Toxocara vitulorum, Ascaris lumbricoides and Necator americanus larval antigen was performed by SDS-PAGE followed by western blotting. A 57 kDa T. canis-specific antibody fraction(TcES-57) was identified by western blotting and labelling with anti-Toxocara antibodies(from experimental rabbits and human patients) and tracing with anti-human or anti-rabbit peroxidase conjugate. No protein fraction of 57 kDa was detected in ES or larval antigens collected from T. canis, T. vitulorum, A. lumbricoides and N. americanus. Using TcES-57, a specific anti-serum was produced in rabbits and a double sandwich ELISA was developed. This test was validated using known seropositive sera from toxocariasis patients, sera from A. lumbricoides or N. americanus patients, and 50 serum samples from cats. These tests revealed that TcES-57 antigen is specific to T. canis infection and does not cross react with sera of other related infections. Thus, ELISA based on TcES-57 antigen was proven to be an effective tool in the diagnosis of toxocariasis and studies on the role of T. canis in the epidemiology of human toxocariasis.
Keywords
Toxocara canis; visceral larva migrans; ELISA; excretory-secretory protein; diagnosis; 57 kDa; Sri Lanka;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Beaver PC, Snyder CH, Carrera GM, Dent JH, Lafferty JW (1952) Chronic eosinophilia due to visceral larva migrans. Report of three cases. Pediatrics 9: 7-19   PUBMED
2 Meghji M, Maizels RM (1986) Biochemical properties of larval excretory-secretory glycoproteins of parasitic nematode Toxocara canis. Mol Biochem Parasitol 18: 155-170   DOI   ScienceOn
3 Rajapakse RPVJ, Lloyd S, Fernando ST (1994) Toxocara vitulorum: maternal transfer of antibodies from buffalo cows (Bubalis bubalis) to calves and levels of infection with T. vitulorum in the calves. Res Vet Sci 57: 81-87   DOI   PUBMED   ScienceOn
4 Wickremasinghe PL, de Wijesundera MS (2001) Ecchymoses- an unusual manifestation of toxocariasis in children. Ceylon Med J 46: 130-131   PUBMED
5 Smith HV, Quinn R, Kusel JR, Girdwood RW (1981) The effect of temperature and antimetabolites on antibody binding to the outer surface of second stage Toxocara canis larvae. Mol Biochem ParasitoI 4(3-4): 183-193   DOI   ScienceOn
6 van Knapen F, van Leusden J, Polderman AM, Franchimont JH (1983) Visceral larva migrans: examination by means of enzyme-linked immunosorbent assay of human sera for antibodies to excretory-secretory antigen of the second stage larvae of Toxocara canis. Z Parasitenkd 63: 113-118   DOI   ScienceOn
7 Kennedy MW, Maizels RM, Meghji M, Young L, Qureshi F, Smith HV (1987) Species-specific carbohydrate epitopes on the secreted and surface antigens of Toxocara cati and Toxocara canis infective larvae. Parasite Immunol 9: 407- 420   DOI   ScienceOn
8 de Savigny DH, Voller A, Woodruff AW (1979) Toxocariasis: serological diagnosis by enzyme-linked immunosorbent assay. J Clin Pathol 32: 284-288   DOI
9 Rajapakse RPVJ, Vasanthathilake VWSM, Lloyd S, Fernando ST (1992) Collection of eggs and hatching and culturing second-stage larvae of Toxocara vitulorum in vitro. J Parasitol 78: 1090-1092   DOI   ScienceOn
10 Maizels RM, de Savigny D, Ogilvie BM (1984) Characterization of surface and excretory-secretory antigens of Toxocara canis infective larvae. Parasite Immunol 6: 23-37   DOI   ScienceOn
11 Speiser F, Gottstein B (1984) A collaborative study on larval excretory-secretary antigens of Toxocara canis for the immunodignosis of human toxocariasis with ELISA. Acta Trop 41: 361-372   PUBMED
12 Gillespie SH, Bidwell D, Voller A, Robertson BD, Maizels RM (1993) Diagnosis of human toxocariasis by antigen capture enzyme-linked immunosorbent assay. J Clin Pathol 46: 551-554   DOI
13 Ash LR, Orihel TC (1987) Parasites: a guide to laboratory procedures and identification. ASCP Press, Chicago
14 Page AP, Richards DT, Lewis JW, Omar HM, Maizels RM (1991) Comparison of isolates and species of Toxocara and Toxascaris by biosynthetic labelling of somatic and ES proteins from infective larvae. Parasitology 103: 451- 464   DOI
15 Radman NE, Archelli SM, Fonrouge RD, del Guardis MV, Linzitto OR (2000) Human toxocarosis. Its seropreva- lence in the City of La Plata. Mem Inst Oswaldo Cruz 95: 281-285   DOI   ScienceOn
16 Starke-Buzetti WA, Ferreira FP (2004) Characterization of excretory / secretory antigen from Toxocara vitulorum larvae. Ann NY Acad Sci 1026: 210-218   DOI   ScienceOn
17 Sugane K, Oshima T (1983) Purification and characterization of excretory-secretory antigen of Toxocara canis larvae. Immunology 50: 113-120   PUBMED
18 Glickman LT, Schantz PM (1981) Epidemiology and patho- enesis of zoonotic toxocariasis. Epidemiol Rev 3: 230- 250   DOI   PUBMED
19 Towbin H, Yaehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350-4354
20 Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of Bacteriophage T4. Nature 227(5259):680-685   DOI   ScienceOn
21 de Savigny DH (1975) In vitro maintenance of Toxocara canis larvae and a simple method for use in serodiagnostic tests for visceral larva migrans. J Parasitol 61: 781-782   DOI   ScienceOn
22 Maizels RM, Robertson BD (1991) Toxocara canis: secreted glycoconjugate antigens in immunobiology and immunodiagnosis. In Parasitic nematodes-antigens, membranes and genes. Kennedy MW (ed.). Taylor and Francis Ltd., London, pp 95-115
23 Jacquier P, Gottstein B, Stingelin Y, Eckert J (1991) Immunodiagnosis of toxocarosis in humans: evaluation of an enzyme-linked immunosorbent assay kit. J Clin Microbiol 29: 1831-1835   PUBMED
24 Glickman LT, Schantz PM, Dombroske R, Cypess R (1978) Evaluation of serodiagnostic tests for visceral larva migrans. Am J Trop Med Hyg 27: 492-498   DOI   PUBMED
25 Lynch NR, Wilkes LK, Hodgen AN, Turner KJ (1988) Specificity of Toxocara ELISA in tropical populations. Parasite Immunol 10: 323-337   DOI   ScienceOn
26 Yamasaki H, Taib R, Watanabe Y, Mak JW, Zasmy N, Araki K, Chooi LPK, Kita K, Aoki T (1998) Molecular characterization of a cDNA encoding an excretory-secretory antigen from Toxocara canis second stage larvae and its application to the immunodiagnosis of human toxocariasis. Parasitol Int 47: 171-181
27 Iddawela RD, Kumarasiri PVR, de Wijesundera MS (2003)A seroepidemiological study of toxocariasis and risk factors for infection in children in Sri Lanka. Southeast Asian J Trop Med Public Health 34: 7-15   PUBMED