Recombinant UL30 antigen-based single serum dilution ELISA for detection of duck viral enteritis

• Published in: Journal of virological methods 2012-11, Vol.185 (2), p.234-238
• Main Authors: Aravind, S., Patil, B.R., Dey, Sohini, Mohan, C. Madhan
• Format: Article
• Language: English
• Subjects: absorbance; Anatid herpesvirus 1; Animals; antibodies; Antibodies, Viral - blood; Antigens, Viral - chemistry; Antigens, Viral - genetics; Biological and medical sciences; blood serum; Cells, Cultured; Chick Embryo; Cloning, Molecular; Duck enteritis virus; ducks; Ducks - virology; Enteritis - diagnosis; Enteritis - veterinary; Enteritis - virology; enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - methods; Enzyme-Linked Immunosorbent Assay - veterinary; equations; Fibroblasts - virology; Fundamental and applied biological sciences. Psychology; genes; Genes, Viral; Mardivirus - chemistry; Mardivirus - genetics; Mardivirus - isolation & purification; Marek Disease - virology; Microbiology; Neutralization Tests; Recombinant antigen; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Regression Analysis; Reproducibility of Results; Sensitivity and Specificity; Single serum dilution ELISA; Techniques used in virology; UL30; viral enteritis; Viral Proteins - chemistry; Viral Proteins - genetics; Viral Proteins - isolation & purification; Virology; Virus Cultivation - methods
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2012.06.027

► A duck enteritis virus was isolated and adapted in chicken embryo fibroblast culture from the State of Kerala, India. ► The UL 30 gene of duck enteritis virus was cloned successfully and expressed from a bacterial prokaryotic system. ► A recombinant UL30 antigen based single serum dilution ELISA was developed for serological diagnosis of duck viral enteritis infection. A recombinant UL30 antigen-based single serum dilution enzyme linked immunosorbent assay (ELISA) was developed to measure specific antibody in the sera of ducks against duck enteritis virus (DEV). The partial UL30 gene of DEV was cloned, expressed, purified and tested for its diagnostic use by designing a single serum dilution enzyme linked immuno-sorbent assay (ELISA). A total of 226 duck sera samples were tested using the assay. A linear relationship was found between the predicted antibody titres at a single working dilution of 1:100 and the corresponding serum titres observed as determined by the standard serial dilution method. Regression analysis was used to determine a standard curve from which an equation was derived which demonstrated this correlation. The equation was then used to convert the corrected absorbance readings of the single working dilution directly into the predicted ELISA antibody titres. The assay proved to be specific, sensitive and accurate as compared to the virus neutralization test with a specificity, sensitivity and accuracy being 96%, 95% and 95% respectively.