Evaluation of the enzyme-linked immunosorbent assay (ELISA) for antibody activity to Anaplasma marginale, Babesia bovis and Babesia bigemina in cattle sera : a thesis /
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| Format: | Thesis Book |
| Language: | English |
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[College Station, Tex.] :
[Texas A&M University],
[1982]
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| Abstract: | ABSTRACT: Comparisons of the efficiency of various buffers for binding of soluble antigens onto solid phase matrices for performance of the ELISA were made. A series of incubation time and temperature combinations for optimum antigen binding was studied. Various antigen preparations of Anaplasma marginale, Babesia bovis and Babesia bigemina were tested for efficiency in detecting antibody activity in bovine sera. Commercially produced horseradish peroxidase labeled rabbit anti-bovine immunoglobulin G (IgG) conjugate was compared to conjugates prepared in the laboratory for efficiency and suitability in the ELISA. Block titrations of antigens, positive and negative sera and conjugates were done to establish optimum dilutions of each for maximum efficiency in the ELISA. Finally, the ELISA was compared with established serological tests for sensitivity and specificity in the detection of antibody activity to A marginale, B bovis and B bigemina. The most suitable combinations of buffers, antigens, conjugate and antigen incubation methods for the 3 parasites were determined as follows: Buffer.... 0.10M carbonate-bicarbonate, pH 9.6. Solid phase matrix.... ELISA "U" bottom microtiter plates. Incubation combination.... 37°C for 16 hours. Conjugate.... Commercial preparation. Antigens.... A marginale -- National Veterinary Services Laboratory (NVSL) whole, sonicated CF antigen. B bovis -- Supernatant from sonicated parasite suspension. B bigemina -- Supernatant from non-sonicated parasite suspension. Test sera.... Dilutions of 1:20. The ELISA was closely comparable to the indirect fluorescent antibody (IFA) test for detection of antibody activity to the 3 parasites. There was also a good correlation between the ELISA and the card test (CT) for detection of A marginale antibody activity. The correlation between the ELISA and the complement fixation (CF) test was not as good. Many sera that were positive ELISA reactors were negative on the CF test. Most of these sera were also positive on the IFA test, which indicated that the ELISA and IFA are probably more sensitive than the CF in detecting low levels of antibody activity. Further studies need to be done to standardize the ELISA accurately, but all indications are that it will be a very useful technique for the serological diagnosis of bovine anaplasmosis and babesiosis. |
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| Item Description: | Typescript (photocopy). Vita. "Major subject: Veterinary Parasitology." "Submitted to the Graduate College of Texas A&M University in partial fulfillment of the requirements for the degree of Master of Science December 1982". Approved as to style and content by: Gerald G. Wagner, Donald E. Corrier, Thomas M. Craig, Ian R. Tizard. |
| Physical Description: | xi, 65 leaves : illustrations ; 29 cm |
| Bibliography: | Includes bibliographical references (leaves 51-53). |