Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1990 Feb;28(2):332–335. doi: 10.1128/jcm.28.2.332-335.1990

Detection of Entamoeba histolytica immunoglobulins G and M to plasma membrane antigen by enzyme-linked immunosorbent assay.

M A Sathar 1, B L Bredenkamp 1, V Gathiram 1, A E Simjee 1, T F Jackson 1
PMCID: PMC269601  PMID: 2107203

Abstract

Sixty-one serum specimens from 22 patients with clinically diagnosed amoebic liver abscess (ALA), 10 hospitalized patients with a variety of diseases other than amoebiasis, 12 normal healthy controls, and 17 subjects from an amoebiasis-endemic area were assayed by enzyme-linked immunosorbent assay (ELISA). The plasma membrane fraction of axenic cultures of Entamoeba histolytica HK9 separated from other subcellular fractions by differential centrifugation was used as the antigen to detect specific immunoglobulin G (IgG) and IgM antibodies. Using a single serum dilution of 1/100 and optical densities at 492 nm of 0.200 and 0.250 as the cutoff values for the IgM and IgG ELISAs, their respective sensitivities in 22 ALA patients were 91% (20 of 22) and 95% (21 of 22). In 22 patients (10 hospitalized and 12 normal healthy controls), the specificities of the IgM and IgG ELISAs were 95% (21 of 22) and 91% (20 of 22), respectively. All five asymptomatic carriers of pathogenic E. histolytica were seropositive by the IgG ELISA and the amoebic gel diffusion test (AGDT). The AGDT was positive for three of six culture-negative controls, while the IgG ELISA was positive for all six. For six asymptomatic carriers of nonpathogenic zymodemes, the AGDT was positive for two, and the IgG ELISA was positive for three. There was an excellent correlation (r = 0.96) between the IgG ELISA and the AGDT. Only one of six culture-negative controls, none of the asymptomatic carriers of pathogenic E. histolytica, and one of six carriers of nonpathogenic E. histolytica were seropositive by the IgM ELISA, thus highlighting the specificity of the IgM ELISA in the diagnosis of ALA. It is believed that the use of plasma membrane fractions has improved the diagnostic potential of the IgM ELISA.

Full text

PDF
332

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aley S. B., Scott W. A., Cohn Z. A. Plasma membrane of Entamoeba histolytica. J Exp Med. 1980 Aug 1;152(2):391–404. doi: 10.1084/jem.152.2.391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bos H. J., Schouten W. J., Noordpool H., Makbin M., Oostburg B. F. A seroepidemiological study of amebiasis in Surinam by the enzyme-linked immunosorbent assay (ELISA). Am J Trop Med Hyg. 1980 May;29(3):358–363. doi: 10.4269/ajtmh.1980.29.358. [DOI] [PubMed] [Google Scholar]
  3. Conradie J. D., Gray R., Mbhele B. E. Serum alphafetoprotein determination by enzyme-linked immunosorbent assay. S Afr Med J. 1980 Jul 26;58(4):169–171. [PubMed] [Google Scholar]
  4. Conradie J. D., Vorster B. J., Kirk R. A simple and rapid method of washing and drying micro-titre plates used in ELISA. J Immunoassay. 1981;2(2):109–116. doi: 10.1080/15321818108056971. [DOI] [PubMed] [Google Scholar]
  5. Gandhi B. M., Irshad M., Acharya S. K., Tandon B. N. Amebic liver abscess and circulating immune complexes of Entamoeba histolytica proteins. Am J Trop Med Hyg. 1988 Nov;39(5):440–444. doi: 10.4269/ajtmh.1988.39.440. [DOI] [PubMed] [Google Scholar]
  6. Gathiram V., Jackson T. F. A longitudinal study of asymptomatic carriers of pathogenic zymodemes of Entamoeba histolytica. S Afr Med J. 1987 Nov 21;72(10):669–672. [PubMed] [Google Scholar]
  7. Goka A. K., Rolston D. D., Mathan V. I., Farthing M. J. Diagnosis of giardiasis by specific IgM antibody enzyme-linked immunosorbent assay. Lancet. 1986 Jul 26;2(8500):184–186. doi: 10.1016/s0140-6736(86)92487-6. [DOI] [PubMed] [Google Scholar]
  8. Jackson T. F., Anderson C. B., Simjee A. E. Serological differentiation between past and present infection in hepatic amoebiasis. Trans R Soc Trop Med Hyg. 1984;78(3):342–345. doi: 10.1016/0035-9203(84)90115-9. [DOI] [PubMed] [Google Scholar]
  9. Jackson T. F., Duursma J., Simjee A. E., Pudifin D. J. Application of the leucocyte adherence inhibition test to the assessment of cell mediated immunity in invasive amoebiasis. Trans R Soc Trop Med Hyg. 1983;77(1):121–125. doi: 10.1016/0035-9203(83)90036-6. [DOI] [PubMed] [Google Scholar]
  10. Jackson T. F., Gathiram V., Simjee A. E. Seroepidemiological study of antibody responses to the zymodemes of Entamoeba histolytica. Lancet. 1985 Mar 30;1(8431):716–719. doi: 10.1016/s0140-6736(85)91262-0. [DOI] [PubMed] [Google Scholar]
  11. Joyce M. P., Ravdin J. I. Antigens of Entamoeba histolytica recognized by immune sera from liver abscess patients. Am J Trop Med Hyg. 1988 Jan;38(1):74–80. doi: 10.4269/ajtmh.1988.38.74. [DOI] [PubMed] [Google Scholar]
  12. Knobloch J., Mannweiler E. Development and persistence of antibodies to Entamoeba histolytica in patients with amebic liver abscess. Analysis of 216 cases. Am J Trop Med Hyg. 1983 Jul;32(4):727–732. doi: 10.4269/ajtmh.1983.32.727. [DOI] [PubMed] [Google Scholar]
  13. Krupp I. M., Powell S. J. Antibody response to invasive amebiasis in Durban, South Africa. Am J Trop Med Hyg. 1971 May;20(3):414–420. doi: 10.4269/ajtmh.1971.20.414. [DOI] [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. Mathews H. M., Moss D. M., Visvesvara G. S. Analysis of antigens from membrane and soluble fractions of Entamoeba histolytica. J Protozool. 1986 Aug;33(3):328–334. doi: 10.1111/j.1550-7408.1986.tb05619.x. [DOI] [PubMed] [Google Scholar]
  16. Meza I., Cázares F., Rosales-Encina J. L., Talamás-Rohana P., Rojkind M. Use of antibodies to characterize a 220-kilodalton surface protein from Entamoeba histolytica. J Infect Dis. 1987 Nov;156(5):798–805. doi: 10.1093/infdis/156.5.798. [DOI] [PubMed] [Google Scholar]
  17. Mohimen A., Maitra T. K., Jalan K. N., Mehra S. A specific solid-phase assay for the detection of immune complexes containing Entamoeba histolytica antigens. J Immunol Methods. 1989 Feb 8;117(1):39–44. doi: 10.1016/0022-1759(89)90116-6. [DOI] [PubMed] [Google Scholar]
  18. Naot Y., Remington J. S. An enzyme-linked immunosorbent assay for detection of IgM antibodies to Toxoplasma gondii: use for diagnosis of acute acquired toxoplasmosis. J Infect Dis. 1980 Nov;142(5):757–766. doi: 10.1093/infdis/142.5.757. [DOI] [PubMed] [Google Scholar]
  19. Petri W. A., Jr, Joyce M. P., Broman J., Smith R. D., Murphy C. F., Ravdin J. I. Recognition of the galactose- or N-acetylgalactosamine-binding lectin of Entamoeba histolytica by human immune sera. Infect Immun. 1987 Oct;55(10):2327–2331. doi: 10.1128/iai.55.10.2327-2331.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Powell S. J., Maddison S. E., Wilmot A. J., Elsdon-Dew R. Amoebic gel-diffusion precipitin-test. Clinical evaluation in amoebic liver abscess. Lancet. 1965 Sep 25;2(7413):602–603. doi: 10.1016/s0140-6736(65)90514-3. [DOI] [PubMed] [Google Scholar]
  21. Ravdin J. I. Pathogenesis of disease caused by Entamoeba histolytica: studies of adherence, secreted toxins, and contact-dependent cytolysis. Rev Infect Dis. 1986 Mar-Apr;8(2):247–260. doi: 10.1093/clinids/8.2.247. [DOI] [PubMed] [Google Scholar]
  22. Robinson G. L. The laboratory diagnosis of human parasitic amoebae. Trans R Soc Trop Med Hyg. 1968;62(2):285–294. doi: 10.1016/0035-9203(68)90170-3. [DOI] [PubMed] [Google Scholar]
  23. Samrejrongroj P., Tharavanij S. Assessment of validity of counterimmunoelectrophoresis and ELISA in the routine diagnosis of amoebiasis. Southeast Asian J Trop Med Public Health. 1985 Sep;16(3):365–370. [PubMed] [Google Scholar]
  24. Sathar M. A., Simjee A. E., Nel J. D., Bredenkamp B. L., Gathiram V., Jackson T. F. Evaluation of an enzyme-linked immunosorbent assay in the serodiagnosis of amoebic liver abscess. S Afr Med J. 1988 Dec 17;74(12):625–628. [PubMed] [Google Scholar]
  25. Torian B. E., Lukehart S. A., Stamm W. E. Use of monoclonal antibodies to identify, characterize, and purify a 96,000-dalton surface antigen of pathogenic Entamoeba histolytica. J Infect Dis. 1987 Aug;156(2):334–343. doi: 10.1093/infdis/156.2.334. [DOI] [PubMed] [Google Scholar]
  26. Yang J., Kennedy M. T. Evaluation of enzyme-linked immunosorbent assay for the serodiagnosis of amebiasis. J Clin Microbiol. 1979 Dec;10(6):778–785. doi: 10.1128/jcm.10.6.778-785.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES