A bio-safe multiple antigenic peptide (MAP) enzyme-linked immunoassay for the detection of antibodies to infectious bron
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ORIGINAL ARTICLE
A bio‑safe multiple antigenic peptide (MAP) enzyme‑linked immunoassay for the detection of antibodies to infectious bronchitis virus in chickens Vikas Verma1 · Vinay G. Joshi1 · Puneet Ranjan1 · Piyush Tomar2 · Rajesh Chhabra3 · N. K. Mahajan2 · Parveen Goel4 Received: 29 April 2020 / Accepted: 31 August 2020 © King Abdulaziz City for Science and Technology 2020
Abstract The objective of the study was to develop a bio-safe synthetic peptide ELISA for the detection of antibodies against the infectious bronchitis virus (IBV) using a novel multiple antigenic peptide approach (MAP). After initial ELISA optimization, diagnostic sensitivity (DSn) and specificity (DSp) for the linear peptides were determined using receiver operator curve (ROC) analysis. The peptide IBVP1 showed 90.44% DSn and 88.64% DSp at ROC cut off 22.8% while IBVP2 showed 88.24% DSn and 85.23% DSp at ROC cut off 23.05%. The multimerization of linear peptides to MAP design resulted in the improvement of the diagnostic efficiency up to 94.85% DSn and 92.05% DSp for IBVM1 with 19.95% cut off. A similar improvement in the performance was also observed with 92.65% DSn and 90.91% DSp for IBVM2 at 20.72% cut off. All the peptides were tested for diagnostic specificity and did not show the cross-reactivity with Newcastle disease virus and infectious bursal disease virus positive serum samples. In addition, repeatability testing for all linear and multimeric peptide showed that the coefficient of variation for intra-assay was within the expected limits, ranging from 2.4 to 10.4% and inter-assay coefficient of variation was ranging from 5.56 to 14.3%. In a nutshell, the present study used predicted B cell epitope, the synthetic peptide in linear and multimeric design for IBV antibody detection. The study also highlights peptide antigen with modified scaffold design could be a safe alternative to whole virion-based ELISA for IBV antibody detection. Keywords Antigenic peptides · Gammacoronavirus of poultry · Peptide scaffold · Peptide diagnostics · Respiratory poultry pathogen
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s13205-020-02422-y) contains supplementary material, which is available to authorized users. * Vinay G. Joshi [email protected] Vikas Verma [email protected]
1
Department of Animal Biotechnology, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana 125 004, India
2
Department of Veterinary Public Health and Epidemiology, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana 125 004, India
3
College Central Laboratory, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana 125 004, India
4
Department of Veterinary Medicine, College of Veterinary Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana 125 004, India
Puneet Ranjan pntrnjn@gmail.
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