DEVELOPMENT OF A PEPTIDE BASED IMMUNOASSAY FOR DETECTION OF ANTIBODIES TO PESTE DES PETITS RUMINANTS VIRUS (PPRV)
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Date
2021
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Abstract
The focus of the present study is to optimize and develop peptide-based immunoassay for the detection of antibodies to PPRV in sheep and goat using synthetic peptides derived from immunoreactive and immunodominant B cell epitopes predicted from Nucleoprotein (N) and Haemagglutinin (H) proteins of PPRV.
Four 8 mer B cell epitopes each for N and H proteins of PPRV namely N1, N2, H1 and H2 were predicted using bioinformatics tools such as abcPRED, BepiPRED 2.0, Phyre 2.0 and IEDB using various indices such as linearity, surface probability, surface accessibility, hydrophilicity, flexibility and antigenicity and were custom synthesized by linking two 8 mer antigenic peptides with GGGS linker and
tested for their immunoreactivity. Based on immunoreactivity the N1, H2 and a cock tail of N1 and H2 was selected as antigen for peptide ELISA.
Serum samples collected at random from sheep and goats vaccinated with live attenuated PPRV vaccine and serum samples collected at specific intervals of 3, 7, 10, 14, 17, 35 days of post vaccination from sheep and goats were used for optimizing the peptide-based immunoassay for PPRV. A total of eighty-eight random serum (Goat = 47, sheep = 41) and Hundred and thirty-five serum samples (Goat = 73, sheep = 62) at specific intervals of vaccination were collected from Post Graduate Research Institute in Animal Sciences (PGRIAS), Kattuppakkam and Instructional Livestock Farm Complex (ILFC), TANUVAS to have a wide range of samples with varying antibody titres.
The serum samples were initially screened with competitive ELISA kit (IVRI, Mukteshwar) before proceeding to peptide ELISA. In competitive ELISA, 106 goat serum samples and 96 sheep serum samples were found to be sero-positive. Fourteen goat serum samples and seven sheep serum sample were shown to be sero negative.