MULTISPECTRAL BACTERIAL IDENTIFICATION

随着探针技术和方法的进步，利用DNA杂交探针进行细菌鉴定已变得越来越复杂和准确。然而，到目前为止，检测荧光标记探针的仪器还不能充分利用这种探针的能力来原位识别高度复杂的基因型混合物。在大量相似细胞中确定单一变异生物体也是有问题的。荧光标记方法最终受到不同光谱“指纹”的数量的限制，这些指纹可以通过用于测量和分类它们的成像系统来区分。我们建议使用一套兼容的荧光团和最近开发的KAIROS仪器以及光谱反褶积和分类算法来增加可以同时区分的荧光标签的数量，从而实现准确的细菌“指纹”。在第一阶段，我们将开发一套这些探针，并证明它们在微观测试目标上的功效。在第二阶段，我们将应用探针库和光谱反褶积软件，通过光谱分选正确识别高度复杂的细胞混合物。这种多光谱细菌鉴定（Mbid）新技术将有利于临床和环境微生物学以及生物技术。建议的商业应用：通过多光谱荧光探针和荧光成像显微分光光度法快速准确地识别细菌指纹，是微生物ID在临床和环境方面的一项使能技术。

Bacterial identification using DNA hybridization probes has become increasingly sophisticated and accurate as probe technology and methods have improved. Up to now, however, instrumentation for detecting fluorescently- labeled probes has not been able to fully exploit the capabilities of such probes to identify high complex mixtures of genotypes in situ. Positive identification of single variant organisms in large populations of similar cells is also problematic. Fluorescent labeling methods are ultimately limited by the number of different spectral 'fingerprints' that can be distinguished by the imaging system that is used to measure and sort them. We propose to use a set of compatible fluorophores and recently developed KAIROS instrumentation and spectral deconvolution and sorting algorithms to increase the number of fluorescent tags that can be simultaneously distinguished, thus enabling accurate 'fingerprinting' of bacteria. In Phase 1 we will develop a set of these probes and demonstrate their efficacy on microscopic test targets. In Phase 2 we will apply the library of probes and the spectral deconvolution software to correctly identify highly complex mixtures of cells in situ by spectral sorting. This new technology for multispectral bacterial identification (Mbid) will benefit clinical and environmental microbiology as well as biotechnology. PROPOSED COMMERCIAL APPLICATION: Rapid and accurate fingerprinting of bacteria by multispectral fluorescent probes and fluorescence imaging microspectrophotometry is an enabling technology for both clinical and environmental aspects of microbial ID.