Chemiluminescence ELISA for the detection of antibodies to bovine leukaemia virus antigens.
A chemiluminescence enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to bovine leukaemia virus antigens (BLV) has been developed. The possibility of using an enhanced chemiluminescence reaction for the determination of adsorbed immunoperoxidase conjugates was studied in this work. The intensity of chemiluminescence depends on both the concentration of reagents and experimental conditions used. The efficiency of the assay is determined by the formation of an immobilized antigen monolayer. A relationship between the quantity of the protein added and adsorbed has been shown. The optimal time and temperature for the antigen-antibody incubation steps have been estimated for each system (3 h at 37 degrees C was chosen as a standard incubation time). A linear dependence of the chemiluminescence intensity and optical density on the concentration of antibodies to the BLV antigens was observed. The detection limit of antibodies in the chemiluminescence ELISA is 2-3 times lower than that in the spectrophotometric one. The results obtained indicate the possibility of using both methods.
Vidziunaité
R
R
Institute of Biochemistry, Vilnius-MTP, Lithuania.
Dikiniené
N
N
Miliukiené
V
V
Mikulskis
P
P
Kulys
J
J
D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000936:Antigen-Antibody Complex; D000956:Antigens, Viral / Q000032:analysis*; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005333:Fetus; D007202:Indicators and Reagents; D007668:Kidney; D001909:Leukemia Virus, Bovine / Q000302:isolation & purification*; D008163:Luminescent Measurements; D051379:Mice / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012756:Sheep; D013053:Spectrophotometry / Q000379:methods
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000936:Antigen-Antibody Complex; D000956:Antigens, Viral; D007202:Indicators and Reagents