Evaluation of antigen-coating procedures of enzyme-linked immunosorbent assay method for detection of trypanosomal antibodies

• Published in: Veterinary parasitology Vol. 90; no. 1; pp. 1 - 13
• Main Authors: Rebeski, D.E, Winger, E.M, Robinson, M.M, Gabler, C.M.G, Dwinger, R.H, Crowther, J.R
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 10-06-2000
• Subjects: Animals; Antibodies, Protozoan - analysis; Antibody; Antigen; Antigens, Protozoan - immunology; Diagnosis; ELISA; Enzyme-Linked Immunosorbent Assay - methods; Enzyme-Linked Immunosorbent Assay - veterinary; Freeze Drying; Polystyrenes; Protozooa; Time Factors; Trypanosoma congolense; Trypanosoma congolense - immunology; Antigen; Diagnosis; Protozooa; Antibody; Trypanosoma congolense; ELISA
• ISSN: 0304-4017; 1873-2550
• DOI: 10.1016/S0304-4017(00)00231-4

Research was undertaken to improve the antigen-coating step of indirect enzyme-linked immunosorbent assay (ELISA) method through the use of polystyrene 96-well plates precoated with antigenically stabile crude trypanosomal antigens. The plates were precoated with antigens, air dried and sealed before being packed in plastic bags with silica gel desiccant packets. Such plates stored at +4 and +37°C provided an assay performance, which was superior to that of plates freshly coated with antigens from a frozen stock. Antigen-precoated plates consistently proved stable after storage up to +50°C for at least 1 year. The accuracy of the assay was not affected, i.e. trypanosomal antibody-positive sera were clearly discriminated from trypanosomal antibody-negative negative sera. In contrast, lyophilized trypanosomal antigens lacked stability on storage at +37°C for longer than 1 month. It was concluded that the routine use of antigen precoated polystyrene plates for the enzyme immunoassay technique will contribute to improved assay robustness at an acceptable diagnostic proficiency. The modified coating procedure will also provide an improved quality assurance and standardization procedure for the assay, which is required to allow the reliable detection of trypanosomal antibodies and comparison of data from different laboratories.