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Indian Journal of Clinical Biochemistry

, Volume 19, Issue 1, pp 88–92 | Cite as

Determination of immunoreactivity ofPlasmodium falciparum antigens, serum dilutions and biomaterials

  • M. M. Mya
  • R. K. Saxena
  • A. Roy
  • D. N. Rao
Article
  • 29 Downloads

Abstract

Immunoreactivity properties of serum dilutions andPlasmodium falciparum malaria antigens were measured and compared by ELISA technique using different ELISA plates to evaluate the role of antigens and serum dilutions for optimum binding. Also effort has been made to see the effect of reaction surface and material i.e. ELISA plates for binding capacity. Serological properties were estimated by ELISA methods for detection of malaria and determination of immunological characteristics. Three Pf antigens (PfAg) i.e. ring infected erythrocyte surface antigen: AR-1 (RESA), histidine-rich protein 2 antigen (HRP-2) and glycophospholipid antigen (grown and developed Pf antigen from PSJ-M strain): GPL1 have been used for serological testing of human blood samples by Enzyme Link Immunosorbant Assay (ELISA). 1∶100, 1∶1000 and 1∶10000 dilutions of Pf positive and negative serum (50 samples in each group) and 1∶1000 dilution of Pf antigens were used to measure immunoreactive properties by ELISA method. Result of PfAg-serum immunoreactivity study showed that GPL1 has the highest degree of immuno binding reactivity compared to other Pf antigens. HRP-2 and RESA antigens showed no significant difference to each other. Study also found that Costar and Fastec ELISA plates have a better Ag−Ab binding capability compared to immulon and Falcon plates at all dilutions of serum. Serum dilution of 1∶100 showed best binding and reactivity with Pf antigens followed by 1∶1000 and 1∶10000 showed lowest reactivity.

Key words

Malaria Serum dilutions GPL1 HRP-2 AR-1 antigens ELISA plates 

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References

  1. 1.
    Soulier, J.P., Prou, O.S. and Dubarry, H. (1985)Plasmodium falciparum antigen detection with monoclonal antibodies. Dev. Biol. Standardization 62, 21–27.Google Scholar
  2. 2.
    Apr, M.E., Evans, C.B. and Taylor, D.W. (1991) Identification ofPlasmodium falciparum histidine rich protein 2 in the plasma of humans with malaria. J. Clin. Microbiol. 29, 1629–1634.Google Scholar
  3. 3.
    Taylor, D.W. and Voller, A. (1993) The development and validation of a simple antigen detection ELISA forPlasmodium falciparum malaria. Trans. R. Soc. Trop. Med. Hyg. 87, 29–31.PubMedCrossRefGoogle Scholar
  4. 4.
    Dietz, R., Perkins, M., Boulos, M., Boulos, M., Luz, F., Reller, B. and Corey, C.R. (1995) The diagnosis ofPlasmodium falciparum using a new antigen detection system. Am. J. Trop. Med. Hyg. 52, 45–49.Google Scholar
  5. 5.
    Gay, F., Traore, B., Zanoni, J., Danis, M. and Gentilini, M. (1994) Evaluation of the QBC system for the diagnosis of malaria. Santi 4, 289–297.Google Scholar
  6. 6.
    Quintana, M., Piper, R., Boling, H.L.et al. (1998) Malaria diagnosis by dipstick assay in a Honduran population with CoendoPlasmodium falciparum andPlasmodium vivax. Am. J. Trop. Med. Hyg. 59, 868–871.PubMedGoogle Scholar
  7. 7.
    Clendennen, T.E., Long, G.W. and Baird, J.K. (1995) QBC and Giemsa stained thick blood films diagnostic performance of laboratory technologists. Trans. R. Soc. Trop. Med. Hyg. 89, 183–184.PubMedCrossRefGoogle Scholar
  8. 8.
    Gay, F., Traore, B., Zanoni, J., Danis, M. and Fribbourg-Blanc, A. (1996) Direct acridine orange fluorescence examination of blood slides compared to current techniques for malaria diagnosis. Trans. R. Soc. Trop. Med. Hyg. 90, 516–518.PubMedCrossRefGoogle Scholar
  9. 9.
    Herrmann, J.E., Hendry, R.M. and Collins, M.F. (1979) Factors involved in enzyme-linked immunoassay of viruses and evaluation of the method for identification of enteroviruses. J. Clin. Microbiol. 10, 210–217.PubMedGoogle Scholar
  10. 10.
    Mya, M.M., Saxena, R.K. and Roy, A. (2002a) Sensitivity and specificity of isolated antigen fromPlasmodium falciparum culture supernatant. Ind. J. Clin. Biochem. 17, 75–82.CrossRefGoogle Scholar
  11. 11.
    Fuchs, S. and Sela, M. (1978) Handbook of experimental immunology. Ed. D.M. Weir (Blackwell Scientific Publications, Oxford). p. 101.Google Scholar
  12. 12.
    Handry, R.M. and Herrmonn, J.E. (1980) Immobilization of antibodies on nylon for use in enzyme-linked immunoassay. J. Immunol. Methods 35, 285–296.CrossRefGoogle Scholar
  13. 13.
    Weetall, H.H. (1970) Preparation and characterization of antigen and antibody adsorbents covalently coupled to an inorganic carrier. Biochem. J. 117, 25–61.Google Scholar
  14. 14.
    Line, W.F. and Becker, M.J. (1975) In immobilized enzymes, antigens, antibodies and peptides, ed. H.H. Weetall (Marcel Dekker, New York) p. 49.Google Scholar
  15. 15.
    Sarkar, M. and Mandal, C. (1985) Immobilization of antibodies on a new solid phase for use in ELISA. J. Immunological Methods 83, 55–60.CrossRefGoogle Scholar
  16. 16.
    Nezlin, R. (2000) Aquantitative approach to the determination of antigen in immune complexes. J. Immunological Methods 237, 1–17.CrossRefGoogle Scholar
  17. 17.
    Mya, M.M., Roy, A., Saxena, R.K. and Roy, K.B. (2002b) Isolation, part characterization, immunogenicity, and ipecificity study ofPlasmodium falciparum culture supernatant. Jpn. J. Infect. Dis. 55, 150–156.PubMedGoogle Scholar
  18. 18.
    Trager, W. and Jensen, J.B. (1976) Human malarial parasites in continuous culture. Science 193, 673–675.PubMedCrossRefGoogle Scholar
  19. 19.
    Folch, J., Lees, M. and Stanley, G.H.S. (1957) A simple method for the isolation and purification of total lipid from animal tissues. J. Biol. Chem. 226, 497–509.PubMedGoogle Scholar
  20. 20.
    Roy, A., Sharma, V.P. and Chauhan, V.S. (1994) The use of peptide ELISA in determinating malaria endemicity. J. Imm. Methods 167, 139–143.CrossRefGoogle Scholar
  21. 21.
    Chiodini, P.L., Hartley, S., Hewett, P.E., Barbara, J.A., Lalloo, K., Bligh, J. and Voller, A. (1997) Evaluation of a malaria antibody ELISA and its value in reducing potential wastage of red cell donations from blood donors exposed to malaria, with a note on a case of transfusiontransmitted malaria. Vox sang 73, 143–148.PubMedCrossRefGoogle Scholar
  22. 22.
    Assal, A., Kauffmann-Lacroix, C., Rodier, M.H., Darde, M.L., Houssay, D. and Jacquemin, J.L. (1999) Comparison de deux techniques de detection des anticorps anti-Plasmodium falciparum: Falciparum-spot IF (Biomerieux et Malaria IgG Celisa (BMD), Resultats Preliminaries. Transfus. Clin. Biol. 6, 119–123.PubMedCrossRefGoogle Scholar
  23. 23.
    Mya, M.M., Saxena, R.K., Bhakat, P. and Roy, A. (2000) Effect of serum dilution in diagnosis of malaria in community. J. Comm. Dis. 32, 28–32.Google Scholar
  24. 24.
    Druilhe, P., Moreno, A., Blanc, C., Brasseur, P.H. and Jacquier, P. (2001) A colorimetricin vitro drug sensitivity assay forPlasmodium falciparum based on a highly sensitive double-site lactate dehydrogenase antigen-capture enzyme-linked immunosorbent assay. Am. J. Trop. Med. Hygn. 64, 233–241.Google Scholar

Copyright information

© Association of Clinical Biochemists of India 2004

Authors and Affiliations

  1. 1.Centre for Biomedical EngineeringIndian Institute of TechnologyNew Delhi
  2. 2.Malaria Research CentreAll India Institute of Medical SciencesNew Delhi
  3. 3.Department of BiochemistryAll India Institute of Medical SciencesNew Delhi

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