ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) MANUFACTURED FOR DIAGNOSIS OF CHICKEN ANAEMIA VIRUS IN BROILER BREEDER FLOCKS

10^th World Congress on HEALTHCARE & TECHNOLOGIES

July 17-18, 2017 | Lisbon, Portugal

Elham O Mahgoub

Qatar University, Qatar

Posters & Accepted Abstracts: J Health Med Informat

Abstract :

Production of indirect enzyme-linked immunosorbent assay (ELISA) to detect serum antibody of chicken anaemia virus (CAV) is described. This test depends on the availability of CAV polyclonal antibodies present in convalescent chicken serum to react with the VP1 antigen and adsorbed to the ELISA plate. In this experiment, VP1 gene is translated to be the capsid protein that holds most the receptor responsible for diagnosis of chicken anaemia virus (CAV). VP1 gene was manufactured to use as coating protein on absorbent face of indirect ELISA. As start, the VP1 gene was inserted into pRSET�B plasmid then transformed into E. coli top10 competent cells. The expressed VP1 protein was then detected using western blot test. Latter after, the VP1 protein been produced in large scale in E. col host. The recombinant VP1 protein was successfully expressed in high cell density. The use of Tangential Flow Filtration (TFF) step was necessary for dialysis and desalting which could increase both the specific activity and the final yield of the purified protein fraction. The protein expressed has been tested as an antigen for detection of antibody to CAV in infected chicken. An enzyme-linked immunosorbent assay (ELISA) of the chicken anaemia virus was prepared for the detection of serum antibody to CAV. sera samples from 60 (positive CAV chicken sera) were screen tested. Statistical analysis methods were applied to measure ELISA specificity, sensitivity, p-value and t-value. In the results and discussions: a band of 50 kDa was showed in western blot test as a proof of the VP1 protein expression. The indirect ELISA specificity was 93.3% and sensitivity was 100%. A t test produced a t-value of 15.805 for the indirect ELISA and revealed a significant difference between CAV-positive serum and CAV-negative serum (p-value of 0.001). For the second variable the t-test yielded a t-value of 5.063, which revealed a significant difference between CAVpositive serum and CAV-negative serum (p-value of 0.015). In conclusions: This indicates that the indirect ELISA approach using VP1 fusion protein has many advantages compared to the commercial indirect ELISA. However, the present study can be a base for the development of ELISA assay for CAV that could reduce the test cost of indirect ELISA method for diagnosis and increase its reliability.