Intact parasite-antigen ELISA test: a new dimension for serodiagnosis of Amoebiasis
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Abstract
An Intact Parasite Antigen ELISA (IPA-ELISA) has been developed for detection of circulating antibodies against Entamoeba. histolytica. Axenically grown trophozoites of E.histolytica (NIH-200) after glutaraldehyde treatment, are used as Intact Parasite Antigen (IPA) as well biological active and imperishable base. Antigen over the surface of treated cells were allowed to interact with the antibody molecules of the test sera. The techniques of Plate, Dot-and IPA-ELISA are compared of their merits for clinical and epidemiological survey of amoebiasis. IPA-ELISA was found to be more sensitive (96.8%) and specific (92.3%) in detecting circulating antibodies compared to Plate-ELISA (sensitivity 90.5%, specificity 84.6%) and Do-ELISA (sensitivity 92.1%, specificity 86.1%). The entire IPA-ELISA test could be completed within 3 hours. Costwise IPA-ELISA is several times lesser than Dot-and Plate-ELISA. This test is most suitable for field and clinical trials for suspected cases of amoebiasis with no skilled hand required for diagnosis.
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Keywords
Amoebiasis, Amoebic liver abscess, Entamoeba histolytica, IPA-ELISA
References
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Dhanalakshmi, S., and Parija, S. C. (2016). Seroprevalence of Entamoeba histolytica from a tertiary care hospital, South India. Tropical Parasitology, 6(1), 78.
Diamond L.S., Harlow D.R.and-Cunnick C.C. (1978)-A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans. R. Soc. Trop. Med. Hyg., 72, 431-432.
Engvall E., Perlmann P. (1971)-Enzyme linked-sorbent assay (ELISA): Quantitative assay of immunoglobulin-G. Immunochemistry, 8, 871-873.
Fierros L.M., Robles M.C.D., Rincon F.E. (1996). Immunodominant Entamoeba histolytica antigen recognized by serum and intestinal antibodies after local or systemic immunization of mice with glutadehyde fixed trophozoites. Life Science, 59, 1283-1295.
Fotedar, R, Stark, D, Beebe, N, Marriott, D, Ellis, J and Harkness, J (2007). Laboratory diagnostic techniques for Entamoeba species. Clin Microbiol Rev. 20, pp. 511–532.
Haghighi A., Rezaeian M. (2005)-Dectection of serum antibody to Entamoeba histolytica in various population samples of amebic infection using an enzyme linked immunosorbent assay. Parasitol. Res., 97(3), 209-212.
Knappik, M., Börner, U., & Jelinek, T. (2005). Sensitivity and specificity of a new commercial enzyme-linked immunoassay kit for detecting Entamoeba histolytica IgG antibodies in serum samples. European Journal of Clinical Microbiology and Infectious Diseases, 24(10), 701-703.
Laemelli U.K. (1970)-Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 277, 680-685.
Leo, M., Haque, R., Kabir, M., Roy, S., Lahlou, R. M., Mondal, D., and Petri, W. A. (2006). Evaluation of Entamoeba histolytica antigen and antibody point-of-care tests for the rapid diagnosis of amebiasis. Journal of clinical microbiology, 44(12), 4569-4571.
Leppard K.N., Totty M., Waterfield E., Harlow J., Jenkins Crawford (1983)-Purification and partial amino acid sequence analysis of the cellular tumour antigen p53 from mouse SV 40 transformed cell. EMBOJ, 2, 1993-1999.
Lopez R.R., Navarro G.F., Valadez-sanchez M., Lopez V.Y., Calva M. J. (1991)-Dot-Enzyme linked immunosorbent assay (Dot-ELISA) of anti Entamoeba histolytica antibodies in human serum and colostrum. Arch Invest. Med. (Mex), 22 (3-4), 249-253.
Mahajan R.C., Sehgal S., Malla N., Ganguly N.K. (1989)-Dot Enzyme linked immunosorbent assay for immuno-diagnosis of ameobiasis. Ind. J.Med.Red., 89, 100-104.
Punthuprapasa P., Thammapalerd N., Chularerk U., Charoenlarp K., Bhaibulaya M. (2001)-Diagnosis of intestinal amoebiasis using salivary IgA antibody detection. Southeast Asian J. Trop. Med. Public Health, 32 (Suppl.) (2) 159-164.
Rosenblatt J.E., Sloan L.M., Bestrom J.E. (1995)-Evaluation of an enzyme linked immunoassay for the detection in serum of antibodies to Entamoeba histolytica. Diag. Microbiol. and Infec. Dis. 22(3):275-278.
Towbin H., Staehelin, T. Gordon (1979)-Elecrophoretic transfer of protein from polyacrylamide gels to nitrocellulose sheets. Proc. Nat. Acad. Sci. USA, 76, 4350-4354.
Trissl D. (1982)-Immunology of Entamoeba histolytica in human and animal hosts. Rev. Infec. Dis., 4, 1154-1160.
Van Doorn H.R., Hofwegen H., Koelewijn R., Gilis H., Peek R., Welsteyn J.C., Van Genderen P.J., Vervoort T., Van Gool T. (2005). Use of rapid dipstick and latex agglutination tests and enzyme-linked immunosorbent assay for serodiagnosis of amebic liver abscess, amebic colitis and Entamoeba histolytica cyst passer. J. Clin. Microbiol., 43(9), 4801-4806.
Van Hal, S. J., Stark, D. J., Fotedar, R., Marriott, D., Ellis, J. T., & Harkness, J. L. (2007). Amoebiasis: current status in Australia. Medical journal of Australia, 186(8), 412.
Walsh J.A. (1986)-Problems in recognition and diagnosis of amoebiasis: estimation of the global magnitude of morbidity and mortality. Rev. Infec. Dis., 8, 228
Wong et al., (2017) also have done similar type of work using crude soluble antigen of E. histolytica
Yamura H., Araki K., Kikuchi K., Itoda I., Totsuka K., Kobayakawa T. (2003), Evaluation of dot-ELISA for serological diagnosis of amoebiasis. J. Infect. Chemother., 9(1), 25-29.
Baveja U.K., Roy T., Kkaur M., Kulpati D.D.S., Agarwal S.k. (1991)- Dot immunobinding assay versus sandwitch ELISA in diagnosis of invasive amoebiasis. Ind. J. Med. Res., 93(A), 161-165.
Dhanalakshmi, S., and Parija, S. C. (2016). Seroprevalence of Entamoeba histolytica from a tertiary care hospital, South India. Tropical Parasitology, 6(1), 78.
Diamond L.S., Harlow D.R.and-Cunnick C.C. (1978)-A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans. R. Soc. Trop. Med. Hyg., 72, 431-432.
Engvall E., Perlmann P. (1971)-Enzyme linked-sorbent assay (ELISA): Quantitative assay of immunoglobulin-G. Immunochemistry, 8, 871-873.
Fierros L.M., Robles M.C.D., Rincon F.E. (1996). Immunodominant Entamoeba histolytica antigen recognized by serum and intestinal antibodies after local or systemic immunization of mice with glutadehyde fixed trophozoites. Life Science, 59, 1283-1295.
Fotedar, R, Stark, D, Beebe, N, Marriott, D, Ellis, J and Harkness, J (2007). Laboratory diagnostic techniques for Entamoeba species. Clin Microbiol Rev. 20, pp. 511–532.
Haghighi A., Rezaeian M. (2005)-Dectection of serum antibody to Entamoeba histolytica in various population samples of amebic infection using an enzyme linked immunosorbent assay. Parasitol. Res., 97(3), 209-212.
Knappik, M., Börner, U., & Jelinek, T. (2005). Sensitivity and specificity of a new commercial enzyme-linked immunoassay kit for detecting Entamoeba histolytica IgG antibodies in serum samples. European Journal of Clinical Microbiology and Infectious Diseases, 24(10), 701-703.
Laemelli U.K. (1970)-Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 277, 680-685.
Leo, M., Haque, R., Kabir, M., Roy, S., Lahlou, R. M., Mondal, D., and Petri, W. A. (2006). Evaluation of Entamoeba histolytica antigen and antibody point-of-care tests for the rapid diagnosis of amebiasis. Journal of clinical microbiology, 44(12), 4569-4571.
Leppard K.N., Totty M., Waterfield E., Harlow J., Jenkins Crawford (1983)-Purification and partial amino acid sequence analysis of the cellular tumour antigen p53 from mouse SV 40 transformed cell. EMBOJ, 2, 1993-1999.
Lopez R.R., Navarro G.F., Valadez-sanchez M., Lopez V.Y., Calva M. J. (1991)-Dot-Enzyme linked immunosorbent assay (Dot-ELISA) of anti Entamoeba histolytica antibodies in human serum and colostrum. Arch Invest. Med. (Mex), 22 (3-4), 249-253.
Mahajan R.C., Sehgal S., Malla N., Ganguly N.K. (1989)-Dot Enzyme linked immunosorbent assay for immuno-diagnosis of ameobiasis. Ind. J.Med.Red., 89, 100-104.
Punthuprapasa P., Thammapalerd N., Chularerk U., Charoenlarp K., Bhaibulaya M. (2001)-Diagnosis of intestinal amoebiasis using salivary IgA antibody detection. Southeast Asian J. Trop. Med. Public Health, 32 (Suppl.) (2) 159-164.
Rosenblatt J.E., Sloan L.M., Bestrom J.E. (1995)-Evaluation of an enzyme linked immunoassay for the detection in serum of antibodies to Entamoeba histolytica. Diag. Microbiol. and Infec. Dis. 22(3):275-278.
Towbin H., Staehelin, T. Gordon (1979)-Elecrophoretic transfer of protein from polyacrylamide gels to nitrocellulose sheets. Proc. Nat. Acad. Sci. USA, 76, 4350-4354.
Trissl D. (1982)-Immunology of Entamoeba histolytica in human and animal hosts. Rev. Infec. Dis., 4, 1154-1160.
Van Doorn H.R., Hofwegen H., Koelewijn R., Gilis H., Peek R., Welsteyn J.C., Van Genderen P.J., Vervoort T., Van Gool T. (2005). Use of rapid dipstick and latex agglutination tests and enzyme-linked immunosorbent assay for serodiagnosis of amebic liver abscess, amebic colitis and Entamoeba histolytica cyst passer. J. Clin. Microbiol., 43(9), 4801-4806.
Van Hal, S. J., Stark, D. J., Fotedar, R., Marriott, D., Ellis, J. T., & Harkness, J. L. (2007). Amoebiasis: current status in Australia. Medical journal of Australia, 186(8), 412.
Walsh J.A. (1986)-Problems in recognition and diagnosis of amoebiasis: estimation of the global magnitude of morbidity and mortality. Rev. Infec. Dis., 8, 228
Wong et al., (2017) also have done similar type of work using crude soluble antigen of E. histolytica
Yamura H., Araki K., Kikuchi K., Itoda I., Totsuka K., Kobayakawa T. (2003), Evaluation of dot-ELISA for serological diagnosis of amoebiasis. J. Infect. Chemother., 9(1), 25-29.
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How to Cite
Intact parasite-antigen ELISA test: a new dimension for serodiagnosis of Amoebiasis. (2018). Journal of Applied and Natural Science, 10(3), 976-980. https://doi.org/10.31018/jans.v10i3.1835
