A Sensitive Immunocapture Real-Time One-Step RT-PCR for Early Detection of Plant Pathogens in Crops

Period of Performance: 01/01/2009 - 12/31/2009


Phase 1 SBIR

Recipient Firm

AC Diagnostics, Inc.
Fayetteville, AR 72701
Principal Investigator


Plant diseases caused by virus infections induce tremendous economical losses for agricultural production each year in the U.S. The increasing international seed trade brings increased potential for the introduction of foreign diseases that could threaten the safety of American agriculture. Early and accurate detection of pant viruses in infested seeds and stock plants is a crucial step to eliminate virus infections and prevent further epidemiological spread among crop plants. Current virus detection is generally performed through the use of biological indexing on indicator plants, enzyme-linked immunosorbent assay (ELISA), or reverse transcription polymerase chain reaction (RT-PCR). In general, bio-indexing is time consuming and labor intensive. Although conventional RT-PCR offers greater sensitivity, it is not cost-effective and difficult to scale-up for high throughput samples. Currently, ELISA remains to be one of the best methods for plant virus detection. A sensitive immunocapture Real-time RT-PCR assay combines the advantages from two widely used virus detection methods, ELISA and RT-PCR to achieve timely and sensitive detection of plant viruses with simple sample preparation by immunocapture technology. This technology provides numerous advantages, including improving sensitivity, reducing contamination risks, eliminating pre- and post-PCR manipulations, and shortening the time needed for large sample diagnosis. This proposal presents the development of a sensitive and early surveillance system for plant viruses in crop plants using this technology. The first objective is to determine the factors affecting the efficient immunocapture of Pepino mosaic virus (PepMV) for Real-time RT-PCR analysis of tomato seed and plant tissues. The second objective is to develop a sensitive and early surveillance system to a panel of 14 tomato viruses using immunocapture Real-time RT-PCR. Once the phase I project is successful, this same strategy could be applied to develop disease surveillance systems for plant viruses in many of the economically important vegetables, fruits, ornamentals and agronomical crops. The goal of this proposal is to develop sensitive, reliable and user-friendly commercial products for routine plant virus detection basin on immunocapture Real-time RT-PCR. This cost-effective and use-friendly detection system will greatly benefit to the growers or companies with limited sources at their diagnostic labs. This novel diagnostic technology based on the immunocapture Real-time RT-PCR will significantly improve plant disease diagnostics by accurate, rapid and cost-effective identification of causal agents in agronomic or specialty crop plants at the earliest possible time relative to manifestation of disease. Therefore, it will improve crop protection resulting in reduced production costs and increased environmental benefits, and enhance protection and safety of the nation's agriculture and food supply.