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

描述（由申请人提供）：最近一项关于炭疽孢子实地测试准确性的政府研究对这些测试的有效性提出了严重怀疑。该报告清楚地表明，迫切需要更严格地选择用于炭疽检测的脱氧核糖核酸（DNA）标记物。拟议中的项目旨在开发一种针对炭疽孢子的更具体的核酸检测方法。PCR引物将被设计用于扩增炭疽菌染色体和质粒中的序列，这些序列可以将炭疽菌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试验的一个挑战。我们将通过探索包括发芽诱导和机械破坏在内的方法组合，开发有效的孢子破坏和核酸提取技术。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传统热循环器所需的1/6时间内进行快速闭管聚合酶链反应（ct-PCR） DNA扩增。此外，系统可以运行6小时的电池供电。在一个试管中结合多个测试的新方法也在开发中，这将超越其他诊断技术平台的能力。封闭管分析将更安全，手持设备将比目前运行的大多数系统更快地提供结果。这种完全自动化的“样本到答案”系统将使最少训练的操作员能够在现场执行复杂的核酸检测。

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.