Advertisement

Detection of Antibodies Against Toxoplasma gondii in Indian Cattle by Recombinant SAG2 Enzyme-Linked Immunosorbent Assay

  • Vikrant Sudan
  • A. K. Tewari
  • Harkirat Singh
Article
  • 6 Downloads

Abstract

Background

A large number of recombinant proteins are tested over a period of time for diagnosis of toxoplasmosis throughout the world. However, such literature is very much limited from an Indian perspective.

Purpose

The aim of the study was to assess the sero prevalence of Toxoplasma gondii in adult cattle from Kerala, India, using recombinant surface antigen 2 (recSAG2) protein.

Methods

An antibody detection recombinant ELISA specific for T gondii was laboratory-standardized using recSAG2 protein. The optimum antigen concentration, serum concentration, and conjugate dilutions were determined by the initial checkerboard titrations. Subsequently, the diagnostic potential of the recombinant protein was assessed with 258 field sera samples cattle and compared with indirect fluorescent antibody test (IFAT).

Results

Among the cattle sera tested, 61.5% showed sero positivity of T. gondii-speciic IgG. When compared to IFAT, the sensitivity of the recSAG2 ELISA was found to be 80.00% with 88.57% specificity at 95% confidence interval with substantial agreement between the tests.

Conclusion

The results of present study have revealed the presence of high seroprevalence of the parasite and, hence, immense public health significance.

Keywords

Cattle IFAT India rec-SAG2-ELISA Toxoplasma gondii 

Notes

Acknowledgements

The authors acknowledge Dr J P Dubey, USDA, Beltsville, USA, for providing the reference sera. The authors are also thankful to the Director, Indian Veterinary Research Institute for providing the facilities as well as to the ICAR for the fellowship awarded to the first author during the perusal of his master’s programme.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Aubert, D., Maine, G.T., Villena, I., Hunt, J.C., Howard, L., Sheu, M., Brojanac, S., Chovan, L.E., Nowlan, S.F. and Pinon, J.M., 2000. Recombinant antigens to detect Toxoplasma gondii specific immunoglobulin G and immunoglobulin M in human sera by enzyme immunoassay. Journal of Clinical Microbiology, 38, 1144–1150.Google Scholar
  2. 2.
    Bachan, M., Deb, A.R., Maharana, B.R., Sudakhar, N.R., Sudan, V., Sarvanan, B.C., Tewari, A.K., 2018. High seroprevalence of Toxoplasma gondii in goats in Jharkhand state of India. Veterinary Parasitology: Regional Studies Reports, 12, 61–68.Google Scholar
  3. 3.
    Balasundaram, M.B., Andavar, R., Palaniswamy, M. and Venkatapathy, N., 2010. Outbreak of acquired ocular toxoplasmosis involving 248 patients. Archives of Ophthalmology 128(1), 28–32.Google Scholar
  4. 4.
    Crawford, J., Lamb, E., Wasmuth, J., Grujic, O., Grigg, M.E. and Boulanger, M.J., 2010. Structural and functional characterization of SporoSAG: A SAG2 related surface antigen from Toxoplasma gondii. Journal of Biological Chemistry, 285, 12063–12070.Google Scholar
  5. 5.
    Dubey, J.P., 1987. Toxoplasmosis in domestic animals in India: present and future. Journal of Veterinary Parasitology, 1, 13–18.Google Scholar
  6. 6.
    Dubey, J.P. and Beattie, C.P., 1988. Toxoplasmosis of Animals and Man. CRC Press, Boca Raton, Florida, 220 pp. 160.Google Scholar
  7. 7.
    Dubey, J.P., Emond, J.P., Desmonts, G. and Anderson, W.R., 1987. Serodiagnosis of postnatally and prenatally induced toxoplasmosis in sheep. American Journal of Veterinary Research, 48, 1239–1243.Google Scholar
  8. 8.
    Dubey, J.P., Somvanshi, R., Jithendran, K.P. and Rao, J.R., 1993. High prevalence of Toxoplasma gondii in goats from Kumaon region of India. Journal of Veterinary Parasitology, 7, 17–21.Google Scholar
  9. 9.
    Fong, M.Y., Lau, Y.L. and Zulqarnain, M., 2008. Characterization of secreted recombinant Toxoplasma gondii surface antigen 2 (SAG2) heterologously expressed by the yeast Pichia pastoris. Biotechnology Letters, 30, 611–618.Google Scholar
  10. 10.
    Jung, C., Lee Cleo, Y-F. and Grigg,M.E., 2004. The SRS superfamily of Toxoplasma surface proteins. International Journal for Parasitology, 34, 285–296.Google Scholar
  11. 11.
    Landis, J.R. and Koch, G.G., 1977. The measurement of observer agreement for categorical data. Biometrics, 33, 159–174.Google Scholar
  12. 12.
    Lau, Y.L. and Fong, M.Y., 2008. Toxoplasma gondii: Serological characterization and immunogenicity of recombinant surface antigen 2 (SAG2) expressed in the yeast Pichia pastoris. Experimental Parasitology, 119, 373–378.Google Scholar
  13. 13.
    Li, S., Galvan, G., Araujo, F.G., Suzuki, Y., Remington, J.S. and Parmley, S., 2000. Serodiagnosis of recently acquired Toxoplasma gondii infection using an enzyme-linked immunosorbent assay with a combination of recombinant antigens. Clinical and Diagnostic Laboratory Immunology,7, 781–787.Google Scholar
  14. 14.
    Palanisamy, M., Madhavan, B., Balasundaram, M.B., Andavar, R. and Venkatapathy, N., 2006. Outbreak of ocular toxoplasmosis in Coimbatore, India. Indian Journal of Ophthalmology, 54(2), 129–131.Google Scholar
  15. 15.
    Parmley, S.F., Sgarlato, G.D., Mark, J., Prince, J.B. and Remington, J.S., 1992. Expression, characterization, and serological reactivity of recombinant surface antigen P22 of Toxoplasma gondii. Journal of Clinical Microbiology, 30, 1127–1133.Google Scholar
  16. 16.
    Prince, J.B., Auer, K.L., Huskinson, J., Parmley, S.F., Araujo, F.G. and Remington, J.S., 1990. Cloning, expression, and cDNA sequence of surface antigen P22 from Toxoplasma gondii. Molecular and Biochemical Parasitology, 43, 97–106.Google Scholar
  17. 17.
    Saeij, J.P.J., Arrizabalaga, G. and Boothroyd, J.C., 2008. A cluster of four surface antigen genes specifically expressed in bradyzoites, SAG2CDXY, plays an important role in Toxoplasma gondii persistence. Infection and Immunity, 76, 2402–2410.Google Scholar
  18. 18.
    Savva, D., Morris, J.C., Johnson, J.D. and Holliman, R.E., 1990. Polymerase chain reaction for detection of Toxoplasma gondii. Journal of Medical Microbiology, 32, 25–31.Google Scholar
  19. 19.
    Sawicka, E.H., Kur, J., Pietkiewicz, H., Holec, L., Gasior, A. and Myjak, P., 2005. Efficient production of the Toxoplasma gondii GRA6, p35 and SAG2 recombinant antigens and their applications in the serodiagnosis of toxoplasmosis. Acta Parasitologica, 50, 249–254.Google Scholar
  20. 20.
    Singh, H., Tewari, A.K., Mishra, A.K., Maharana, B.R., Sudan, V., Raina, O.K. and Rao, J.R., 2014. Detection of antibodies to Toxoplasma gondii in domesticated ruminants by recombinant truncated SAG2 enzyme-linked immunosorbent assay. Tropical Animal Health and Production,  https://doi.org/10.1007/s11250-014-0703-5.
  21. 21.
    Sudan, V., Tewari, A.K., Singh, H., 2015. Serodiagnosis of Toxoplasma gondii infection in bovines from Kerala, India using a recombinant surface antigen 1 ELISA. Biologicals, 43(4), 250–255.Google Scholar
  22. 22.
    USDHEW (U.S. Department of Health, Education and Welfare) Manual, 1976. A procedural guide for the performance of the serology of toxoplasmosis. Centre for Disease Control, Atlanta.Google Scholar
  23. 23.
    Vastava, P.B., Pradhan, S., Jha, S., Prasad, K.N., Kumar, S. and Gupta, R.K., 2002. MRI features of toxoplasma encephalitis in the immunocompetent host: a report of two cases. Neuroradiology, 44,834–838.Google Scholar
  24. 24.
    Velmurugan, G.V., Tewari, A.K., Rao, J.R., Baidya, S., Kumar,M.U. and Mishra, A.K., 2008. High-level expression of SAG1 and GRA7 gene of Toxoplasma gondii (Izatnagar isolate) and their application in serodiagnosis of goat toxoplasmosis. Veterinary Parasitology, 154, 185–192.Google Scholar
  25. 25.
    Voller, A. and O’Neill, P., 1971. Immunofluorescence methods suitable for large scale application to malaria. Bulletin of World Health Organization, 45, 524–529.Google Scholar

Copyright information

© Witold Stefański Institute of Parasitology, Polish Academy of Sciences 2019

Authors and Affiliations

  • Vikrant Sudan
    • 1
    • 2
  • A. K. Tewari
    • 1
    • 3
  • Harkirat Singh
    • 1
    • 4
  1. 1.Indian Veterinary Research Institute (IVRI)BareillyIndia
  2. 2.Department of Parasitology, College of Veterinary Sciences and Animal HusbandryUP Pandit Deen Dayal Upadhyaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU)MathuraIndia
  3. 3.Division of ParasitologyIndian Veterinary Research Institute (IVRI)BareillyIndia
  4. 4.Department of Veterinary Parasitology, College of Veterinary ScienceGADVASULudhianaIndia

Personalised recommendations