MGB Eclipse Probe Detection of Category A Organisms

A 类生物的 MGB Eclipse 探针检测

• 批准号: 6689127
• 负责人: IRINA A AFONINA
• 金额: $ 10万
• 依托单位: ELITECHGROUP, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-15 至 2003-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6689127
• 关键词: Bacillus anthracis; Clostridium botulinum; DNA primers; Yersinia pestis; biohazard detection; biomarker; biotechnology; bioterrorism /chemical warfare; fluorescent dye /probe; nucleic acid hybridization; nucleic acid probes; polymerase chain reaction; single nucleotide polymorphism; technology /technique development

DESCRIPTION (provided by investigator): Fluorogenic minor groove binder (MGB) probes containing an MGB-quencher at the 5'-end and a fluorophore at the 3'-end have been recently reported. These probes fluoresce on hybridization to the complementary targets. The 5'-MGB-quencher group prevents 5' - nuclease digestion by Taq polymerase during homogeneous amplification. The 5'-MGB-quencher-oligonucleotide-fluors can be used as probes in general nucleic acid and SNP detection assays. We propose to use a new software prediction program to develop MGB Eclipse probe assays to detect Clostridium botulinum, Bacillus anthracis and Yersinia pestis. The designed probes and primers will be optimized with use of three modified bases to improve probe performance of AT- and GC-rich sequences. In the Phase 1 effort, we will design and optimize the primers and probes and demonstrate function of the MGB Eclipse probe assays on a set of known target samples. The specificity of the probes will be evaluated against panels of closely related non-pathogenic organisms. In Phase II, we will expand the number of Category A organisms and improve throughput by multiplexing. In addition, commercial prototype MGB Eclipse(TM) probe assays will be developed for each of these organisms.

描述（由研究者提供）：最近报道了在5 '端含有MGB-猝灭剂和在3'端含有荧光团的荧光小沟结合剂（MGB）探针。这些探针在与互补靶标杂交时发荧光。5 '-MG-猝灭剂基团在均相扩增期间防止Taq聚合酶对5' -核酸酶的消化。5 ′-MG-猝灭剂-荧光素可用作一般核酸和SNP检测测定中的探针。我们建议使用一个新的软件预测程序，开发MGB Eclipse探针检测肉毒梭菌，炭疽杆菌和鼠疫耶尔森氏菌。设计的探针和引物将使用三个修饰的碱基进行优化，以提高AT和GC富集序列的探针性能。在第1阶段的工作中，我们将设计和优化引物和探针，并证明MGB Eclipse探针检测试剂盒对一组已知目标样本的功能。将针对密切相关的非致病性微生物组评价探针的特异性。在第二阶段，我们将扩大A类微生物的数量，并通过多路复用提高通量。此外，将为这些微生物中的每一种开发商业原型MGB Eclipse（TM）探针测定。