MGB Eclipse Probe Detection of Category A 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槽粘结剂（MGB）探针。这些探针在杂交到互补靶标上时发出荧光。在均匀扩增过程中，5'- mgb -淬灭基团阻止Taq聚合酶对5'-核酸酶的消化。5'- mgb -淬灭剂-寡核苷酸荧光可作为一般核酸检测和SNP检测的探针。我们建议使用一个新的软件预测程序来开发MGB Eclipse探针检测肉毒杆菌、炭疽芽孢杆菌和鼠疫杆菌。设计的探针和引物将使用三个修饰的碱基进行优化，以提高富含AT和gc的序列的探针性能。在第一阶段的工作中，我们将设计和优化引物和探针，并在一组已知的目标样品上演示MGB Eclipse探针检测的功能。探针的特异性将在密切相关的非致病性生物体中进行评估。在第二阶段，我们将扩大A类生物的数量，并通过多路复用提高产量。此外，商业原型MGB Eclipse（TM）探针分析将为每一个这些生物开发。

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.