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Authors: Verma, Vikas
Advisor: Ganeshrao, (Joshi Vinay
Title: “Study on peptide based enzyme linked immune-sorbent assay for the diagnosis of infectious bronchiti
Publisher: LUVAS
Language: en
Type: Thesis
Agrotags: null
Keywords: Infectious Bronchitis Virus (IBV), Peptide ELISA
Abstract: Infectious bronchitis virus (IBV), is an important pathogen of poultry which is the prototype of subfamily Coronavirinae, family Coronaviridae, order Nidovirales. It primarily affects respiratory tract but some other clinical manifestations like enteric, nephritic and reproductive disorders are also observedInfectious bronchitis virus (IBV) continues to be one of the most important poultry pathogens worldwide.The commercially available ELISA kits forIBV antibody detection are mostly based on the whole virion and few serological tests based on non-structural proteins of IBV have been developed.Therefore, in the present study we tried to develop an easy alternative indirect ELISA for detection of IBV antibody using antigenic epitope of S1 spike glycoprotein of IBV as coating antigen instead of using whole virus as coating antigen with additional advantages like, lower probability of cross-reactivity of peptides. Two novel peptides, SP1 and SP2, two previously published peptides i.e. IBV-1 and IBV-2, and their MAPs i.e. MAP-1 and MAP-2 were synthesized using solid phase peptide synthesis (SPPS). Two novel peptides i.e. SP1 and SP2, were used as coating antigen in peptide ELISA and their results were compared with two commercial ELISA kits for IB i.e. IDEXX and BIO-CHECK.The diagnostic sensitivity (DSN), specificity (DSP) for SP1 were 95.70% and 92.70% respectively when compared with IDEXX in 87 field sera, were 91.90% and 85.70% respectively, WHEN compared with BIO-CHECK in same 87 field sera samples. For SP2, DSN and DSP were 93.80% and 93.70% respectively, when compared with IDEXX and 96.30% and 93.30% respectively, when compared with BIO-CHECK in same 87 field serum samples. Furthermore, we also performed nano blot assay and flow through assay using all the synthesized peptides as coating antigen and found appreciable LSPR (Localized Surface Plasmon Resonance) response showing the positive reaction against commercial IBV antibodies. The LSPR can be further enhanced using enhancer solutions. The results presented in this study indicate that the developed peptide ELISA has the potentialto serve as a rapid, reliable and cost-effective method for IBV antibody detection.
Subject: Animal Biotechnology
Theme: “Study on peptide based enzyme linked immune-sorbent assay for the diagnosis of infectious bronchiti
These Type: M.V.Sc.
Issue Date: 2018
Appears in Collections:Thesis

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