Killed avian influenza virus (AIV) vaccines have been used to control H5N1 infections in countries where the virus is endemic. Distinguishing vaccinated from naturally infected birds (DIVA) in such situations however, has become a major challenge. Recently, we introduced the recombinant ectodomain of the M2 protein (M2e) of H5N1 subtype as a novel tool for an ELISA based DIVA test. Despite being antigenic in natural infection the monomer form of the M2e used in ELISA had limited antigenicity and consequently poor diagnostic capability. To address this shortcoming, we evaluated the use of four tandem copies of M2e (tM2e) for increased efficiency of M2e antibody detection. The tM2e gene of H5N1 strain from Indonesia (A/Indonesia/CDC540/2006) was cloned into a pMAL- p4x expression vector and expressed in @entity526 as a recombinant @entity11231 . Both of these, M2e and tM2e antigens reacted with sera obtained from @entity1573 following live @entity281 but not with sera from vaccinated birds. A significantly stronger M2e antibody reaction was observed with the tM2e compared to M2e antigen. Western blotting also supported the superiority of tM2e over M2e in detection of specific M2e antibodies against live @entity281 . Results from this study demonstrate that M2e tetramer is a better antigen than single M2e and could be more suitable for an ELISA based DIVA test.

Multimeric Recombinant M2e Protein-Based ELISA: A Significant Improvement in Differentiating Avian Influenza Infected XXXX from Vaccinated Ones.

@entity1573 :: ('9031', 'Species') :: ['chickens'] @entity281 :: ('MESH:D007239', 'Disease') :: ['H5N1 infection'] @entity526 :: ('562', 'Species') :: ['E.coli'] @entity11231 :: ('MESH:C563602', 'Disease') :: ['tM2e-MBP or M2e-MBP proteins']
@entity1573 :: (9031,Species) :: ['Chickens']
