Hand-Held System to Detect Anthrax Spores in Bio-defense

描述（由申请人提供）：政府最近对炭疽孢子现场测试的准确性进行了一项研究，对这些测试的有效性产生了严重怀疑。 这份报告清楚地表明，迫切需要更严格地选择用于炭疽测试的脱氧核糖核酸（DNA）标记。 拟议的项目旨在开发一种更具体的炭疽孢子核酸测试。 PCR引物将被设计用于扩增炭疽细菌染色体和质粒中的序列，这些序列可以将炭疽与相关物种区分开来。从炭疽孢子中快速提取核酸也是现场测试的一个挑战。 我们将开发有效的孢子破碎和核酸提取技术，通过探索方法的组合，包括发芽诱导和机械破碎。 然后将核酸提取和扩增方案纳入我们专有的试管实验室（Liat（TM））平台，以开发适合现场测试的手持设备。 Liat系统已被证明能够在常规热循环仪所需时间的1/6内进行快速闭管聚合酶链反应（ct-PCR）DNA扩增。 此外，该系统可以在电池电源下运行6小时。 将联合收割机多项测试结合在一个试管中的新方法也在开发中，这种方法将超过其他诊断技术平台的能力。 一个封闭的试管分析将更安全地实施和手持设备将提供结果更快地比大多数系统目前在运作。 这一全自动的“采样到应答”系统将使经过最低限度培训的操作员能够在现场进行复杂的核酸检测。

DESCRIPTION (provided by applicant): A recent government study on the accuracy of field tests for anthrax spores has cast serious doubt on the validity of these tests. This report clearly indicates the urgent need for a more rigorous selection of deoxyribonucleic (DNA) markers for anthrax testing. The proposed project aims to develop a more specific nucleic acid test for anthrax spores. PCR primers will be designed to amplify sequences in the anthrax bacterial chromosome and plasmids that can differentiate anthrax from related species. Rapid extraction of nucleic acids from anthrax spores is also a challenge for field tests. We will develop efficient spore disruption and nucleic acid extraction techniques by exploring a combination of approaches, including germination induction and mechanical disruption. The nucleic acid extraction and amplification protocols will then be incorporated into our proprietary lab-in-a-tube (Liat (TM)) platform to develop a handheld device that is suitable for field tests. The Liat system has been shown to be capable of performing rapid close-tube polymerase chain reaction (ct-PCR) DNA amplifications in 1/6 the time required by conventional thermal cyclers. Moreover, the system can be operated for 6 hours on battery power. Novel approaches to combine multiple tests in a single tube that will surpass the capability of other diagnostic technology platforms are also under development. A closed-tube assay will be safer to implement and the hand-held device will deliver results more quickly than most of the systems currently in operation. This fully automated "sample-to-answer" system will enable minimally trained operators to perform sophisticated nucleic acid tests in the field.