Specific monoclonal antibodies to Strongyloides stercoralis: a potential diagnostic reagent for strongyloidiasis.
In this study, specific hybridomas secreting monoclonal antibodies (MAb) to antigen of Strongyloides stercoralis filariform larvae were produced. Specific epitopes targeted by the MAb were protein in nature and located in situ in the internal content of the filariform larvae of the parasite but not in the esophagus. The MAb reacted to the homologous antigen in an indirect ELISA but did not reveal any reaction to the SDS-PAGE separated-homologous antigen in a Western blot analysis (WB) suggesting a conformational epitope specificity. The MAb were of IgG1 isotype which is the isotype known to have high affinity to this epitope so they were used in a dot-ELISA to detect the antigen of the parasite. The assay could detect the epitopes in 78 ng or more of the crude filariform larval extract but did not reveal any positive result when applied to detect antigen in stool samples of parasitologically confirmed strongyloidiasis patients. The negative antigen test results can be explained as follows. Either the MAb were filariform stage-specific and thus did not recognize the rhabditiform larval antigen mainly contained in the patient's stool or the amounts of antigen in the stool samples were too small and/or unevenly dispersed. In the latter instance, the MAb developed in this study would have a diagnostic potential if used in an immunological test design where more volume of fresh stool sample could be accommodated in the test, e.g. a sandwich plate ELISA.
Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
Piyanan
P
Taweethavonsawat
Wanpen
W
Chaicumpa
Urai
U
Chaisri
Usa
U
Chuenbal
Yuwaporn
Y
Sakolvaree
Pramuan
P
Tapchaisri
Thitima
T
Wongsaroj
D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology*; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000947:Antigens, Helminth / Q000276:immunology*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000939:Epitopes / Q000276:immunology; D005243:Feces / Q000469:parasitology; D006801:Humans; D006825:Hybridomas; D007814:Larva / Q000276:immunology; D051379:Mice; D012680:Sensitivity and Specificity; D017171:Strongyloides stercoralis / Q000276:immunology*; D013322:Strongyloidiasis / Q000175:diagnosis* / Q000276:immunology
D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D000939:Epitopes