The isolation, by differential antibody screening, of Echinococcus multilocularis antigen gene clones with potential for immunodiagnosis.
A lambda gt11 cDNA expression library consisting of a million recombinant clones has been constructed from Echinococcus multilocularis protoscolex mRNA. Differential screening of the library with pools of E. multilocularis and Echinococcus granulosus human infection sera has revealed 13 potentially immunodiagnostic clones. On the basis of plaque immunoassays and lysogen characteristics, two of these clones, designated EM2 and EM4, have been further characterised. The recombinant fusion-peptides have been purified and their potential as immunodiagnostic reagents has been assessed by immunoblotting and, in the case of one fusion-peptide (EM4), by enzyme-linked immunosorbent assay (ELISA). Furthermore, the native parasite antigens coded for by these clones have been identified. EM2 corresponds to a 70 kDa protein and epitopes coded for by EM4 have been found on three antigens of 62, 49 and 44 kDa. The native antigens of both clones are present in the protoscolex and those corresponding to EM4 appear to be excreted/secreted products. They are not recognised in ELISA by a variety of human parasitic infection sera other than sera taken from patients infected with E. multilocularis. Nevertheless, the native antigens for both clones are present in E. granulosus protoscoleces and Taenia solium cysticerci. These antigens are not detectable in E. granulosus cyst fluid, and this may, in part, explain the lack of immune response to them in human E. granulosus and T. solium infections.
Hemmings
L
L
Department of Pure and Applied Biology, Imperial College of Science and Technology, London, U.K.
McManus
D P
DP
D000818:Animals; D000909:Antibodies, Helminth / Q000302:isolation & purification; D000947:Antigens, Helminth / Q000235:genetics* / Q000276:immunology; D010582:Bacteriophage lambda / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods; D004443:Echinococcosis / Q000175:diagnosis / Q000276:immunology / Q000469:parasitology; D004446:Echinococcus / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D005796:Genes; D006801:Humans; D007118:Immunoassay / Q000379:methods; D015151:Immunoblotting; D007159:Immunologic Tests; D008242:Lysogeny; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D010948:Viral Plaque Assay
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D011993:Recombinant Fusion Proteins