Comparison of Different Immunosorbent Assays for Citrus Tristeza Virus (CTV) Using CTV Specific Monoclonal and Polyclonal Antibodies

Abstract

Production of monoclonal antibodies specific to CTV has improved CTV detection and allowed the use of different immunoenzymatic techniques. In this study the indirect (I), DAS, and double antibody sandwich indirect (DASI) ELISA was compared for ability to detect CTV in plant extracts and in purified preparations. Twenty-four variations of ELISA were tested using CTV-specific monoclonal and polyclonal antibodies, anti-immunoglobulins, protein A and the biotinlavidin system. Alkaline phosphatase and peroxidase-labeled enzymes were also compared. The DASI ELISAvariations with CTV-specific polyclonal antibodies used as coating antibodies and monoclonal antibodies used as the intermediate antibody detected 0.1 ng/ml CTV protein. The limit of detection for other variations was less than 100 ng/ml CTV protein.