Development of a multi-epitope antigen of S protein-based ELISA for antibodies detection against infectious bronchitis virus
| Autor: | Wen-qiao Fan, Hongning Wang, Peng-wei Xu, Hai-peng Cao, Anyun Zhang, Xin Yang, Bing-cun Ma, Meng-die Ding |
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| Rok vydání: | 2015 |
| Předmět: |
Infectious bronchitis virus
Enzyme-Linked Immunosorbent Assay Applied Microbiology and Biotechnology Biochemistry Antibodies Analytical Chemistry Epitopes Western blot Antigen medicine Animals Molecular Biology Gene Antigens Viral biology medicine.diagnostic_test Organic Chemistry General Medicine Multi epitope Virology Molecular biology Primary and secondary antibodies Fusion protein Spike Glycoprotein Coronavirus biology.protein Antibody Chickens Biotechnology |
| Zdroj: | Bioscience, biotechnology, and biochemistry. 79(8) |
| ISSN: | 1347-6947 |
| Popis: | An indirect enzyme-linked immunosorbent assay (ELISA) method based on a novel multi-epitope antigen of S protein (SE) was developed for antibodies detection against infectious bronchitis virus (IBV). The multi-epitope antigen SE protein was designed by arranging three S gene fragments (166–247 aa, S1 gene; 501–515 aa, S1 gene; 8–30 aa, S2 gene) in tandem. It was identified to be approximately 32 kDa as a His-tagged fusion protein and can bind IBV positive serum by western blot analysis. The conditions of the SE-ELISA method were optimized. The optimal concentration of the coating antigen SE was 3.689 μg/mL and the dilution of the primary antibodies was identified as 1:1000 using a checkerboard titration. The cut-off OD450 value was established at 0.332. The relative sensitivity and specificity between the SE-ELISA and IDEXX ELISA kit were 92.38 and 89.83%, respectively, with an accuracy of 91.46%. This assay is sensitive and specific for detection of antibodies against IBV. |
| Databáze: | OpenAIRE |
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