Development of A New Instrument to Identify Microbial Genome

描述（由申请人提供）：在应对生物恐怖袭击或传染病爆发时，快速识别和追踪病原菌的来源是必要的。脉冲场凝胶电泳（PFGE）指纹图谱经常用于这些任务，特别是用于追踪细菌的起源。然而，当使用延长的分离时间来解析长DNA分子时，短片段条带在PFGE中丢失，从而降低检测特异性，因为短片段的长度提供了额外的基因型信息，这对于区分两个相似的基因组可能是至关重要的。PFGE的另一个缺点是电泳速度太慢。在快速（24小时）PFGE方法中，电泳需要14-18小时。此外，实验室内和实验室间的重现性 PFGE也需要改进。尽管已经探索了多种方法用于DNA分离，例如熵阱和DNA棱镜，但迄今为止还没有一种被实际使用。我们最近发现了一种新的和有效的技术，以解决广泛的大小范围的DNA分子。在此应用中，我们建议构建一种新的仪器来证明这种新技术的原理，用于高速和准确的微生物鉴定。

DESCRIPTION (provided by applicant): Rapid identification and tracing the origin of pathogenic bacteria are imperative in response to a bioterrorist attack or an infectious disease outbreak. Pulsed-Field Gel Electrophoresis (PFGE) fingerprinting is frequently used for these tasks, particularly for tracing bacterium origins. However, the short fragment bands are lost in PFGE when extended separation times are used to resolve the long DNA molecules, thus reduce the detection specificity, because the lengths of the short fragments provide additional genotype information that could be critical to discriminate two similar genomes. Another drawback of PFGE is that the electrophoresis is too slow. In a rapid (24-hour) PFGE method the electrophoresis takes 14-18 hours. Additionally, the intra- and inter-laboratory reproducibility of PFGE also needs to be improved. Although diverse approaches such as entropic traps and DNA prisms have been explored for DNA separations, none have thus far been practically utilized. We have recently discovered a new and efficient technique to resolve broad size ranges of DNA molecules. In this application we propose to construct a novel instrument to demonstrate the proof-of-principle of this new technique for high-speed and accurate microbial identification.