Antibody pairs for the detection of EV71 and CA16.

Abstract

An immunochromatographic lateral flow assay (LFA) was developed for rapid detection and diagnosis of hand-foot-and-mouth disease (HFMD) using monoclonal antibodies (MAbs) against a pair of peptides, VP1 and VP2 of EV71, the causative agent of HFMD. VP1 and VP2 are capsid proteins that are exposed on the virion surface. They are responsible for host-receptor binding and immune response. Supernatants containing MAbs raised against VP1 and VP2 peptides from hybridoma clones were screened by GenScript using enzyme linked immune sorbent assay (ELISA). Twelve MAbs were tested positive and shipped by GenScript with alphanumeric codes for easy identification. These MAbs were screened to determine binding specificity against EV71 virion using indirect immunofluorescence. Four MAbs were able to recognise EV71 virion. They are, Anti-VP1 5B10B10, Anti-VP1 5B10C8, Anti-VP2 4G2G5 and Anti-VP2 2D9E5. A pair of lyophilised MAbs, Anti-VP2 4G2G5 and Anti-VP1 4D9E7 were also given by GenScript. Only Anti-VP2 4G2G5 can recognise EV71 using indirect immunofluorescence. Anti-VP2 4G2G5 were also tested positive against EV71 and VP2 proteins in LFA set up. Acris, a commercially available Anti-EV71 MAb can also recognise EV71, VP1 and VP2 proteins in LFA and it is more sensitive than lyophilised Anti-VP2 4G2G5 MAbs.