ESTABLISHMENT OF AN ELISPOT ASSAY FOR DETECTION OF ACTIVE HUMORAL IMMUNITY AGAINST CLASSICAL SWINE FEVER VIRUS IN PIGS

摘要

Classical swine fever is one of the major infectious diseases that cause serious economic loss to swine industry worldwide. The ineffective control program in Thailand is partly related to the lack of information regarding to the active immunologicalstatus in pigs, and therefore, improper vaccination protocol. Although the routinely used technique, neutralizing peroxidase linked assay, can be used to evaluate the level of serum neutralizing liter, this assay does not differentiate the actively produced antibody from the passively transferred maternal antibody in the young pigs. An ELISA-SPOT or ELISPOT assay has been reportedly used for studies of the immune responses in pig in several models. The assay has been used to demonstrate the induction ofactive humoral immunity in several lymphoid organs and tissues. In order to assess active humoral immunity in pigs following immunization, in this study we established an ELISPOT assay for detection of classical swine fever virus (CSFV)-specific antibody secreting cells (ASC) from porcine peripheral blood mononuclear cells (PBMC).