Improved ELISA for the detection of adenovirus antigen in faeces extracts by the biotin/streptavidin interaction.
Two enzyme-linked immunosorbent assays for detection of adenovirus antigen in faecal extracts have been established. A conjugate of rabbit anti-(human) adenovirus immunoglobulin and horseradish peroxidase (HRP) prepared by means of a hetero-bifunctional reagent, N-succinimidyl-3-(2-pyridyldithio) propionate (SPDP), was used in the conventional ELISA, and a biotin-labelled antibody in combination with a streptavidin/peroxidase complex in the other (BS-ELISA). A collection of 60 faecal extracts, in which adenovirus had been demonstrated by immunosorbent electron microscopy (ISEM) in 29 samples, was selected and examined by ELISA and BS-ELISA. The presence of adenovirus was demonstrated in 38 (63%) of the samples by BS-ELISA compared to 34 (58%) by ELISA, showing the advantage of the biotin/streptavidin system for setting up ELISA assays. Comparison of the three different techniques showed that the biotin/streptavidin (BS-ELISA) is the most sensitive assay for detection of adenovirus antigen in faecal samples.
D000260:Adenoviruses, Human / Q000276:immunology* / Q000302:isolation & purification / Q000648:ultrastructure; D000956:Antigens, Viral / Q000032:analysis*; D001426:Bacterial Proteins; D001710:Biotin; D004797:Enzyme-Linked Immunosorbent Assay; D005243:Feces / Q000382:microbiology*; D006801:Humans; D007074:Immunoglobulin G; D008854:Microscopy, Electron; D019809:Streptavidin
D000956:Antigens, Viral; D001426:Bacterial Proteins; D007074:Immunoglobulin G; D001710:Biotin; D019809:Streptavidin