Evaluation of a 70 kDa Excreted/Secreted Coproantigen Immunoassay for the Detection of Toxocara canis in Dogs
-
Ana Cristina González-Morales
,
Zinnia Judith Molina-Garza
,
Ricardo Gomez-Flores
,
Juan José Zárate-Ramos
,
Lucio Galaviz-Silva
Abstract
Background: We aimed to develop a sandwich ELISA, using polyclonal antibodies against excretory/secretory (E/S) antigens specific to coproantigens present in Toxocara canis-positive dogs.
Methods: Antibodies were produced at Biological Sciences School, Autonomous University of Nuevo León, México in 2023 by immunization of rabbits with antigenic extracts from in vitro cultures of T. canis larvae. Assays were performed on 100 stool samples from pet dogs, measuring sensitivity, specificity, and cross-reactivity against other parasitic infections.
Results: High values of sensitivity (100%), specificity (90.9%), and positive (93.47%) and negative (95.45%) predictive values were obtained, respectively. We obtained an E/S protein with a molecular weight of 70 kDa, which showed high sensitivity and specificity by ELISA, but it presented cross-reactivity against Ancylostoma caninum and Strongyloides stercoralis.
Conclusion: Additional studies are necessary to increase the specificity percentage since this assay demonstrated significant potential as a useful and inexpensive diagnostic tool for immunodiagnostic in dog feces.
1. Mohamad S, Azmi NC, Noordin R. Development and evaluation of a sensitive and specific assay for the diagnosis of human toxocariasis using three recombinant antigens (TES-26, TES-30USM and TES-120). J Clin Microbiol. 2009;47(6):1712 –1717.
2. Apt BW. Most frequent parasite infections and their management. Rev Med Clin Condes. 2014;25(3):485 –528.
3. Yamasaki H, Araki K, Lim PK, et al. Development of a highly specific recombinant second-stage larval excretory-secretory antigen from Toxocara canis for the immunodiagnosis of human toxocariasis. J Clin Microbiol. 2000;38(4):1409-1413.
4. Naderbandi M, Zibaei M, Haniloo A, et al. Larva migrans in BALB/c mice experimentally infected with Toxocara cati ensured by PCR assay. BMC Vet Res. 2022;18:257.
5. Durán ML, García LG, Menchén DA, et al. Parasitosis with dermatological manifestati-ons. Medicine-Accredited Continuing Medical Education Program. 2022;13(59):3493–3503.
6. Olave AM, Mesa JA, Botero JH, et al. Production and evaluation of the Toxocara canis recombinant antigen TES-30 for the immunodiagnosis of toxocariasis. Biomedica. 2016;36(1):39–51.
7. Zibaei M, Sadjjadi SM, Sarkari B, Uga S. Evaluation of Toxocara cati excretory-secretory larval antigens in serodiagnosis of human toxocariasis. J Clin Lab Anal. 2016; 30(3): 248-53.
8. Dolz G, Guzmán GN, Bermúdez FR, et al. Detection of anti-Toxocara spp antibodies in canines from a peri-urban community in the province of Heredia, Costa Rica. Ciencias Veterinarias. 2020; 38(1):28-42.
9. Durant JF, Irenge LM, Fogt WR, et al. Duplex quantitative real-time PCR assay for the detection and discrimination of Toxocara canis and Toxocara cati (Nematoda, Ascaridoidea) eggs in soil and feces samples. Parasit Vectors. 2012:5:288.
10. Ezema KU, Malgwi SA, Zango MK, et al. Gastrointestinal parasites of dogs (Canis familiaris) in Maiduguri, Borno State, Northeastern Nigeria: Risk factors and zoonotic implications for human health. Vet World. 2019;12(7):1150–1153.
11. Nieves A, Ortega B, Martinez M, et al. Standardization of ELISA technique for the immunological diagnosis of human toxocariasis. Bol Malariol Salud Ambient. 2012;52(1):21-32.
12. Jin Y, Shen C, Huh S, et al. Serodiagnosis of toxocariasis by ELISA using crude antigen of Toxocara canis larvae. Korean J Parasitol. 2013;51(4):433–439.
13. De Savigny DH. In vitro maintenance of Toxocara canis larvae and a simple method to produce Toxocara ES antigen for use in serodiagnostic tests for visceral larva migrans. J Parasitol. 1975;61(4):781-782.
14. Zibaei M, Uga S. Modified method to enhanced recovery of Toxocara cati larvae for the purposes of diagnostic and therapeutic. Exp Parasitol. 2016; 169:107-110.
15. Bradford M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-254.
16. Jara LM, Rodriguez M, Altamirano F, et al. Development and validation of a copro-enzyme-linked immunosorbent assay sandwich for detection of Echinococcus granulosus-soluble membrane antigens in dogs. Am J Trop Med Hyg. 2019;100(2): 330–335.
17. Elsemore DA, Geng J, Cote J, et al. Enzyme-linked immunosorbent assays for coproantigen detection of Ancylostoma caninum and Toxocara canis in dogs and Toxocara cati in cats. J Vet Diagn Invest. 2017;29(5):645–653.
18. Santos LM, Cerqueira MP, Gaboardi GC, et al. Evaluation of glycosylated TES from Toxocara canis produced in Pichia pastoris for the immunodiagnosis of human toxocariasis. Braz Arch Biol Technol. 2020;63.
19. Del Águila C, Cuellar C, Guillén JL. Toxocara canis excretory/secretory antigen: polyclonal and larvicidal monoclonal antibody recognition profiles. Parasite Immunol. 1988;10(2):237-241.
20. Voller A, Bartlett A, Bidwell DE. Enzyme immunoassays for parasitic diseases. Trans R Soc Trop Med Hyg. 1976;70 (2):98-106.
21. Espinoza Y, Huapaya P, Suárez R, et al. Standardization of the ELISA technique for the diagnosis of human Toxocariasis. An Fac Med Med. 2003;64(1):7-12.
22. Iglesias SA, Olivos K, Failoc V. Toxocariasis, a disease that still deserves our attention. Experience in Medicine Magazine of the Lambayeque Regional Hospital. 2018;4(4):159-160.
23. Vélez L, Barrera RK, Almaráz RD, et al. Potential risk of zoonotic parasites present in canine feces in Puerto Escondido, Oaxaca. Salud Publica Mex. 2014;56(6):625-630.
24. Olave J, García P, Martínez V, et al. Prevalence of gastrointestinal helminths in dogs from the Health Service of Tulancingo, Hidalgo. Abanico Vet. 2019;9:1-10.
25. Moura MQ, Macedo RP, Terto WD, et al. Detection of Toxocara canis DNA in tissues of experimentally infected mice. Acta Trop. 2018; 187:51–56.
26. Vizcaychipi KA, Rinas M, Irazu L, et al. Neotropical zoonotic parasites in bush dogs (Speothos venaticus) from Upper Paraná Atlantic Forests in Misiones Argentina. Vector Borne Zoonotic Dis. 2016;16(10):664–672.
27. Morales OL. Identification of Toxocara canis antigens using the Western blot technique in experimentally infected rabbits. Doctoral Thesis. 1999.
28. Gazzinelli PH, Nutman TB. Helminth parasites and immune regulation. F1000Res. 2018; 7:F1000 Faculty Rev-1685.
29. Roldán WH, Rubinsky EG. Immunodiagnosis of human toxocariasis. In Serological diagnosis of certain human, animal and plant diseases. Intech Open. 2012.
30. Ávila, HG, Risso MG, Ruybal P, et al. Development of a low-cost copro-LAMP assay for the simultaneous coprodetection of Toxocara canis and Toxocara cati. Parasitology. 2021;148 (7):819-826.
31. Stangogiannis DE, Marval H, Moreno MM, et al. Experimental infection in mice with ocular larva migrans. Arch Soc Esp Oftalmol. 2007; 82(2):89-94.
2. Apt BW. Most frequent parasite infections and their management. Rev Med Clin Condes. 2014;25(3):485 –528.
3. Yamasaki H, Araki K, Lim PK, et al. Development of a highly specific recombinant second-stage larval excretory-secretory antigen from Toxocara canis for the immunodiagnosis of human toxocariasis. J Clin Microbiol. 2000;38(4):1409-1413.
4. Naderbandi M, Zibaei M, Haniloo A, et al. Larva migrans in BALB/c mice experimentally infected with Toxocara cati ensured by PCR assay. BMC Vet Res. 2022;18:257.
5. Durán ML, García LG, Menchén DA, et al. Parasitosis with dermatological manifestati-ons. Medicine-Accredited Continuing Medical Education Program. 2022;13(59):3493–3503.
6. Olave AM, Mesa JA, Botero JH, et al. Production and evaluation of the Toxocara canis recombinant antigen TES-30 for the immunodiagnosis of toxocariasis. Biomedica. 2016;36(1):39–51.
7. Zibaei M, Sadjjadi SM, Sarkari B, Uga S. Evaluation of Toxocara cati excretory-secretory larval antigens in serodiagnosis of human toxocariasis. J Clin Lab Anal. 2016; 30(3): 248-53.
8. Dolz G, Guzmán GN, Bermúdez FR, et al. Detection of anti-Toxocara spp antibodies in canines from a peri-urban community in the province of Heredia, Costa Rica. Ciencias Veterinarias. 2020; 38(1):28-42.
9. Durant JF, Irenge LM, Fogt WR, et al. Duplex quantitative real-time PCR assay for the detection and discrimination of Toxocara canis and Toxocara cati (Nematoda, Ascaridoidea) eggs in soil and feces samples. Parasit Vectors. 2012:5:288.
10. Ezema KU, Malgwi SA, Zango MK, et al. Gastrointestinal parasites of dogs (Canis familiaris) in Maiduguri, Borno State, Northeastern Nigeria: Risk factors and zoonotic implications for human health. Vet World. 2019;12(7):1150–1153.
11. Nieves A, Ortega B, Martinez M, et al. Standardization of ELISA technique for the immunological diagnosis of human toxocariasis. Bol Malariol Salud Ambient. 2012;52(1):21-32.
12. Jin Y, Shen C, Huh S, et al. Serodiagnosis of toxocariasis by ELISA using crude antigen of Toxocara canis larvae. Korean J Parasitol. 2013;51(4):433–439.
13. De Savigny DH. In vitro maintenance of Toxocara canis larvae and a simple method to produce Toxocara ES antigen for use in serodiagnostic tests for visceral larva migrans. J Parasitol. 1975;61(4):781-782.
14. Zibaei M, Uga S. Modified method to enhanced recovery of Toxocara cati larvae for the purposes of diagnostic and therapeutic. Exp Parasitol. 2016; 169:107-110.
15. Bradford M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-254.
16. Jara LM, Rodriguez M, Altamirano F, et al. Development and validation of a copro-enzyme-linked immunosorbent assay sandwich for detection of Echinococcus granulosus-soluble membrane antigens in dogs. Am J Trop Med Hyg. 2019;100(2): 330–335.
17. Elsemore DA, Geng J, Cote J, et al. Enzyme-linked immunosorbent assays for coproantigen detection of Ancylostoma caninum and Toxocara canis in dogs and Toxocara cati in cats. J Vet Diagn Invest. 2017;29(5):645–653.
18. Santos LM, Cerqueira MP, Gaboardi GC, et al. Evaluation of glycosylated TES from Toxocara canis produced in Pichia pastoris for the immunodiagnosis of human toxocariasis. Braz Arch Biol Technol. 2020;63.
19. Del Águila C, Cuellar C, Guillén JL. Toxocara canis excretory/secretory antigen: polyclonal and larvicidal monoclonal antibody recognition profiles. Parasite Immunol. 1988;10(2):237-241.
20. Voller A, Bartlett A, Bidwell DE. Enzyme immunoassays for parasitic diseases. Trans R Soc Trop Med Hyg. 1976;70 (2):98-106.
21. Espinoza Y, Huapaya P, Suárez R, et al. Standardization of the ELISA technique for the diagnosis of human Toxocariasis. An Fac Med Med. 2003;64(1):7-12.
22. Iglesias SA, Olivos K, Failoc V. Toxocariasis, a disease that still deserves our attention. Experience in Medicine Magazine of the Lambayeque Regional Hospital. 2018;4(4):159-160.
23. Vélez L, Barrera RK, Almaráz RD, et al. Potential risk of zoonotic parasites present in canine feces in Puerto Escondido, Oaxaca. Salud Publica Mex. 2014;56(6):625-630.
24. Olave J, García P, Martínez V, et al. Prevalence of gastrointestinal helminths in dogs from the Health Service of Tulancingo, Hidalgo. Abanico Vet. 2019;9:1-10.
25. Moura MQ, Macedo RP, Terto WD, et al. Detection of Toxocara canis DNA in tissues of experimentally infected mice. Acta Trop. 2018; 187:51–56.
26. Vizcaychipi KA, Rinas M, Irazu L, et al. Neotropical zoonotic parasites in bush dogs (Speothos venaticus) from Upper Paraná Atlantic Forests in Misiones Argentina. Vector Borne Zoonotic Dis. 2016;16(10):664–672.
27. Morales OL. Identification of Toxocara canis antigens using the Western blot technique in experimentally infected rabbits. Doctoral Thesis. 1999.
28. Gazzinelli PH, Nutman TB. Helminth parasites and immune regulation. F1000Res. 2018; 7:F1000 Faculty Rev-1685.
29. Roldán WH, Rubinsky EG. Immunodiagnosis of human toxocariasis. In Serological diagnosis of certain human, animal and plant diseases. Intech Open. 2012.
30. Ávila, HG, Risso MG, Ruybal P, et al. Development of a low-cost copro-LAMP assay for the simultaneous coprodetection of Toxocara canis and Toxocara cati. Parasitology. 2021;148 (7):819-826.
31. Stangogiannis DE, Marval H, Moreno MM, et al. Experimental infection in mice with ocular larva migrans. Arch Soc Esp Oftalmol. 2007; 82(2):89-94.
| Files | ||
| Issue | Vol 19 No 4 (2024) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijpa.v19i4.17162 | |
| Keywords | ||
| Helminthology Toxocara canis Excreted-secreted anti-gens Immunodiagnostics Toxocariasis | ||
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How to Cite
1.
González-Morales AC, Molina-Garza ZJ, Gomez-Flores R, Zárate-Ramos JJ, Galaviz-Silva L. Evaluation of a 70 kDa Excreted/Secreted Coproantigen Immunoassay for the Detection of Toxocara canis in Dogs. Iran J Parasitol. 2024;19(4):418-427.
