Development of a Blocking ELISA Based on a Monoclonal Antibody Against a Predominant Epitope in Non-structural Protein 3B2 of Foot-and-mouth Disease Virus for Differentiating Infected from Vaccinated Animals

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2014 Nov 5

PMID 25369323

Citations 7

Authors

Yuanfang Fu

Zengjun Lu

Pinghua Li

Yimei Cao

Pu Sun

Meina Tian

Na Wang

Huifang Bao

Xingwen Bai

Dong Li

Yingli Chen

Zaixin Liu

Affiliations

Soon will be listed here.

Abstract

A monoclonal antibody (McAb) against non-structural protein (NSP) 3B of foot-mouth-disease virus (FMDV) (3B4B1) was generated and shown to recognize a conserved epitope spanning amino acids 24-32 of 3B (GPYAGPMER) by peptide screening ELISA. This epitope was further shown to be a unique and predominant B cell epitope in 3B2, as sera from animals infected with different serotypes of FMDV blocked the ability of McAb 3B4B1 to bind to NSP 2C3AB. Also, a polyclonal antibody against NSP 2C was produced in a rabbit vaccinated with 2C epitope regions expressed in E. coli. Using McAb 3B4B1 and the 2C polyclonal antibody, a solid-phase blocking ELISA (SPB-ELISA) was developed for the detection of antibodies against NSP 2C3AB to distinguish FMDV-infected from vaccinated animals (DIVA test). The parameters for this SPB-ELISA were established by screening panels of sera of different origins. Serum samples with a percent inhibition (PI) greater than or equal to 46% were considered to be from infected animals, and a PI lower than 46% was considered to indicate a non-infected animal. This test showed a similar performance as the commercially available PrioCHECK NS ELISA. This is the first description of the conserved and predominant GPYAGPMER epitope of 3B and also the first report of a DIVA test for FMDV NSP 3B based on a McAb against this epitope.

Citing Articles

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