utilization of monoclonal antibodies in immunocapture RT-PCR and dot blotting immunobinding assays for the detection of Cucumber mosaic virus

ABSTRACT

Reliable plant diagnostic techniques are necessary for good crop management. Occasionally, ELISA may not give a definitive result, requiring confirmation using different diagnostic methods. Immunocapture reverse transcription PCR [IC-RT-PCR] requires the direct confirmation of an ELISA result in a simple, cost-effective manner. We found that the use of captured antigens as a template in IC-RT-PCR can eliminate the need for traditional extraction methods and reduce the number of steps involved, costs incurred, and sampling variations. Experiments showed that a reliability of pathogens can be detected using ELISA in the IC-RT-PCR process. To date, a rapid assay for diagnosis of CMV has not been developed, and there is a critical need for a method that can be employed in both laboratory and field. Dot immunobinding assays [DIBA] are useful alternatives to microtitre plate enzyme-linked immunosorbent assay [ELISA]. It has the additional advantages of simplicity, and is completed quickly in the field or the laboratory on large numbers of samples, in a short time. The method proposed provides a sensitive quantitative technique for dot-immunobinding assay