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.

这个小企业创新研究项目是开发一种名为Amplidisks的创新技术，有助于检测种子中的细菌性植物病原体。健康的种子对美国和全世界的农业都非常重要。在一批种子中，只有少数受感染的种子（0.01%）会导致疾病流行和严重的作物损失。由于种子批中受感染种子的频率通常非常低，因此需要高灵敏度的种子检测方法。目前可用的测定使用液体种子提取物的小等分试样（0.01-0.1%），并且可能导致假阴性结果。该测定将通过使用新的富集技术实现数百倍更大的种子提取物等分试样的采样，其中靶生物体的数量在Ampli-disk中增加，并通过实时PCR检测和确认。第一阶段工作的目的是显示该技术用于检测番茄种子中的Clavibacter michiganensis pv michiganensis的可行性。II期项目的后续工作将验证该程序，并开发包含Ampli-disk和含有所有PCR成分的干珠的商业试剂盒。 该项目的商业应用将在农业领域。基于所提出的技术的检测试剂盒的开发和销售预计将导致消除患病的种子批次和食品材料。更健康的种子可能会提高作物产量，改善消费者的食品安全。