Development of a monoclonal antibody-based enzyme-linked immunosorbent assay for the detection of live propagules of Trichoderma harzianum in a peat-bran medium

Abstract

Mouse monoclonal antibodies (MAbs) and mouse and rabbit (polyclonal) antisera were raised against a Trichoderma harzianum isolate T95. A cell line, Th.HD3 3, raised from splenocytes from a mouse immunized with lyophilized mycelium and Quil A adjuvant, produced MAbs of the immunoglobulin sub-class IgG 2a that were specific recognizing, by ELISA, antigens from a number of Trichoderma species and species from the related genus Hypocrea but not from a range of other soil-borne fungi. These MAbs were used to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of live propagules of Trichoderma harzianum in a peat-bran medium. Heat, protease and periodate treatment of the T95 antigen and binding on Western blots indicate that the Th.HD3 3 MAbs recognized a protein epitope of ca M r 18,500 which forms part of a larger glycoprotein molecule.