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

SBIR 第一阶段：利用新型富集技术结合自动实时 PCR 定量检测种子中的细菌病原体

• 批准号: 0320498
• 负责人: Parm Randhawa
• 金额: $ 10万
• 依托单位: California Seed and Plant Lab., Inc.
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2003-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0320498&HistoricalAwards=false
• 关键词: SBIR; Phase; Quantitative; Detection; Bacterial

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.

这个小型企业创新研究项目是开发一种称为Amplidisks的创新，该创新将有助于检测种子中的细菌植物病原体。健康种子对美国和世界各地的农业非常重要。种子批次中只有少数受感染的种子（0.01％）会导致疾病流行和明显的作物丧失。由于种子地块中感染种子的频率通常非常低，因此需要高度敏感的种子测试方法。当前可用的测定方法使用液态种子提取物的小等分试样（0.01-0.1％），并可能导致虚假负面结果。该测定法可以通过使用一种新颖的富集技术来对种子提取物的几百倍取样，在这种富集技术中，在放大器中，目标生物的数量增加，并通过实时PCR检测并确认并确认。第一阶段工作的目的是展示该技术在番茄种子中检测密歇根州PV密歇根州的可行性。 II阶段项目的后续项目将验证该程序并开发包含所有PCR成分的放大器盘和干珠的商业套件。 该项目的商业应用将在农业领域。根据提出的技术，预计测试套件的开发和营销将导致消除患病的种子土地和食物材料。更健康的种子可能会导致农作物产量提高，并改善消费者的食品安全。