SBIR Phase I: Quantitative Detection of Bacterial Pathogens in Seeds by Use of a Novel Enrichment Technique Coupled with Automated Real-Time PCR

California Seed And Plant Lab., Inc., Elverta CA

Investigators

Abstract

This Small Business Innovation Research project is to develop an innovation called Amplidisks that would assist in the detection of bacterial plant pathogens in seeds. Healthy seeds are very important to agriculture in the United States and throughout the world. Just a few infected seeds (0.01%) in a seed lot can result in a disease epidemic and significant crop loss. Since the frequency of infected seeds in a seed lot is generally very low, highly sensitive seed testing methods are needed. Currently available assays use small aliquots (0.01-0.1%) of a liquid seed extract, and can lead to false negative results. This assay would enable sampling of several hundred times larger aliquots of seed extract by using a novel enrichment technique, where the number of target organisms are increased in an Ampli-disk and detected and confirmed by real-time PCR. The objective of the Phase I work is to show the feasibility of this technique for detection of Clavibacter michiganensis pv michiganensis in tomato seeds. The follow on Phase II project will validate the procedure and develop commercial kits containing Ampli-disks and dry beads with all PCR ingredients. The commercial application of this project will be in the area of agriculture. The development and marketing of test kits based on the the proposed technique is expected to result in the elimination of diseased seed lots and food materials. Healthier seeds would likely lead to increased crop yields and to improved food safety for the consumer.