Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1991 Sep;29(9):1836–1841. doi: 10.1128/jcm.29.9.1836-1841.1991

Rapid, low-technology field- and laboratory-applicable enzyme-linked immunosorbent assays for immunodiagnosis of Schistosoma mansoni.

C L Rossi 1, V C Tsang 1, J B Pilcher 1
PMCID: PMC270220  PMID: 1774304

Abstract

Simple and rapid polystyrene- and nitrocellulose-based enzyme-linked immunosorbent assays were developed for detecting antibodies against adult Schistosoma mansoni microsomal antigens. The polystyrene test uses the Nunc Immuno Stick System. A single dilution of the antibody source being tested, the conjugate, and the substrate (3,3',5,5'-tetramethylbenzidine) are placed in tubes. Dried, antigen-coated polystyrene sticks are then exposed to the reagents by immersion. Once the sticks are sensitized, an entire assay can be completed in 8 min. Positive reactions result in a rich blue color in the substrate tube and can be distinguished with the naked eye. In the nitrocellulose-based test, a nitrocellulose sheet with antigen drawn in a line by pen is cut to produce identical strips. The ligand-binding steps and washings are performed in the troughs of incubation trays. The exposure times required for a single dilution of the antibody source being tested, the conjugate, and the substrate (3,3'-diaminobenzidine) are 5 min, 5 min, and 7.5 min, respectively. Once sensitized strips are available, an entire assay can be run in 50 min. Both techniques can assay serum or whole blood. The characteristics of polystyrene- and nitrocellulose-based techniques allow them to be used successfully in field studies and in minimally equipped laboratories.

Full text

PDF
1836

Selected References

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

  1. Aeppli R. E., Bargetzi M. J., Binz H. Diagnostic screening of multiple antigen-antibody reactions in a new single assay on nitrocellulose: the line immunobinding assay (LIBA). J Immunol Methods. 1989 Jun 2;120(1):93–98. doi: 10.1016/0022-1759(89)90293-7. [DOI] [PubMed] [Google Scholar]
  2. Brand J. A., Tsang V. C. A rapid immunoblot assay (western blot) to detect specific antibodies for human immunodeficiency virus, Schistosoma mansoni, and Taenia solium (Cysticercosis). J Immunoassay. 1989;10(2-3):237–255. doi: 10.1080/01971528908053239. [DOI] [PubMed] [Google Scholar]
  3. Brand J. A., Tsang V. C., Zhou W., Shukla S. B. Comparison of particulate 3,3',5,5'-tetramethylbenzidine and 3,3'-diaminobenzidine as chromogenic substrates for immunoblot. Biotechniques. 1990 Jan;8(1):58–60. [PubMed] [Google Scholar]
  4. Hancock K., Tsang V. C. Development and optimization of the FAST-ELISA for detecting antibodies to Schistosoma mansoni. J Immunol Methods. 1986 Sep 27;92(2):167–176. doi: 10.1016/0022-1759(86)90162-6. [DOI] [PubMed] [Google Scholar]
  5. Raoult D., Dasch G. A. The line blot: an immunoassay for monoclonal and other antibodies. Its application to the serotyping of gram-negative bacteria. J Immunol Methods. 1989 Dec 20;125(1-2):57–65. doi: 10.1016/0022-1759(89)90078-1. [DOI] [PubMed] [Google Scholar]
  6. Tsang V. C., Hancock K., Maddison S. E., Beatty A. L., Moss D. M. Demonstration of species-specific and cross-reactive components of the adult microsomal antigens from Schistosoma mansoni and S. japonicum (MAMA and JAMA). J Immunol. 1984 May;132(5):2607–2613. [PubMed] [Google Scholar]
  7. Tsang V. C., Hancock K., Maddison S. E. Quantitative capacities of glutaraldehyde and sodium m-periodate coupled peroxidase-anti-human IgG conjugates in enzyme-linked immunoassays. J Immunol Methods. 1984 May 11;70(1):91–100. doi: 10.1016/0022-1759(84)90393-4. [DOI] [PubMed] [Google Scholar]
  8. Tsang V. C., Tsang K. R., Hancock K., Kelly M. A., Wilson B. C., Maddison S. E. Schistosoma mansoni adult microsomal antigens, a serologic reagent. I. Systematic fractionation, quantitation, and characterization of antigenic components. J Immunol. 1983 Mar;130(3):1359–1365. [PubMed] [Google Scholar]
  9. Tsang V. C., Wilson B. C., Peralta J. M. Quantitative, single-tube, kinetic-dependent enzyme-linked immunosorbent assay (k-ELISA). Methods Enzymol. 1983;92:391–403. doi: 10.1016/0076-6879(83)92033-5. [DOI] [PubMed] [Google Scholar]

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

RESOURCES