Application of a recombinant Echinococcus multilocularis antigen in an enzyme-linked immunosorbent assay for immunodiagnosis of human alveolar echinococcosis

Mol Biochem Parasitol. 1989 Sep;36(2):151-9. doi: 10.1016/0166-6851(89)90187-4.

Abstract

A highly antigenic polypeptide fragment of the recombinant Echinococcus multilocularis antigen II/3 was produced in Escherichia coli and purified for application in enzyme-linked immunosorbent assay (ELISA). The antigen II/3-encoding 1.0 kb DNA sequence was reduced by sonication into smaller DNA fragments which were subsequently cloned into lambda gt11. Three clones could be isolated from the sublibrary, all synthesizing a recombinant antigen as a stable beta-galactosidase fusion protein. In a further step, the 0.6-kb insert from one positive clone was subcloned into the plasmid pAR 3038, which directed efficient synthesis of the antigen fused to only 11 amino acids from the N-terminus of the phage T7 major capsid protein. The plasmid-encoded antigen (antigen II/3-10) was purified from a bacterial cell extract and then tested in an ELISA. Using sera from 88 patients with an E. multilocularis-infection, a high diagnostic sensitivity of 90% was demonstrated. Investigation of sera from 220 patients with various helminthic infections showed a specificity of 99%, suggesting the suitability of the antigen II/3-10 as an immunodiagnostic tool.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Helminth / analysis*
  • Antigens, Helminth* / biosynthesis
  • Antigens, Helminth* / genetics
  • Antigens, Helminth* / isolation & purification
  • Blotting, Western
  • Echinococcosis / diagnosis*
  • Echinococcosis / immunology
  • Echinococcus / immunology*
  • Enzyme-Linked Immunosorbent Assay*
  • Escherichia coli / genetics
  • Genetic Vectors
  • Humans
  • Plasmids
  • Recombinant Fusion Proteins / immunology
  • Transformation, Bacterial

Substances

  • Antibodies, Helminth
  • Antigens, Helminth
  • Recombinant Fusion Proteins