Avidin-biotin amplification procedures for the detection of human terminal deoxynucleotidyl transferase in cell smears.
The authors have developed fluorescent avidin and avidin-peroxidase assays to detect human terminal deoxynucleotidyl transferase (TdT) in cell smears. These assays used specifically purified rabbit anti-calf TdT antibody, biotinylated goat anti-rabbit IgG antibody, and either fluorescein isothiocyanate-avidin or avidin-biotinylated horseradish peroxidase complex. The use of phosphate-buffered saline rather than TRIS-buffered saline or borate-buffered saline was essential to obtain optimal results in the fluorescent avidin assay. Fixation of cells with either 0.5% paraformaldehyde or 10% neutral buffered formalin was found to be superior to either no fixation or fixation with cold methanol or cold 0.1% glutaraldehyde in ethanol. The sensitivities of the fluorescent avidin and avidin-peroxidase assays were shown to be identical, based on staining intensities and percentages of positive cells when both assays were performed on cells from 14 patients with acute lymphoblastic leukemia (ALL), and 5 patients with lymphoblastic lymphoma (LL).
D000741:Anemia, Aplastic / Q000201:enzymology; D000818:Animals; D001360:Avidin; D001710:Biotin; D004253:DNA Nucleotidylexotransferase / Q000097:blood*; D004254:DNA Nucleotidyltransferases / Q000097:blood*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000276:immunology; D007945:Leukemia, Lymphoid / Q000201:enzymology*; D007951:Leukemia, Myeloid / Q000201:enzymology; D011817:Rabbits
D007074:Immunoglobulin G; D001360:Avidin; D001710:Biotin; D004254:DNA Nucleotidyltransferases; D004253:DNA Nucleotidylexotransferase