Rapid NAT System for Point-of-care SARS Diagnosis

描述（由申请人提供）：本研究申请的目标是基于我们专有的试管实验室（Liat TM）平台开发一种快速、易于使用、集成的严重急性呼吸综合征（SARS）核酸检测。Liat系统能够在单个封闭管系统中集成样品制备、多重核酸扩增和实时定量。最近的SARS疫情表明，地方一级需要更灵敏、快速和自动化的核酸检测能力，以更好地诊断患者并提高检疫效果。完成后，拟议的系统将能够在不到1小时的时间内采集原始血浆或痰液样本，并在护理点进行全自动样本到答案SARS相关冠状病毒（SCV）核酸检测。在满足这些要求的情况下，该系统将在生物防御和民用医疗保健方面具有广泛的应用。为了实现I期目标，将设计并验证几种SCV特异性小沟结合（MGB）-TaqMan探针。使用硅胶磁珠或核酸杂交方法进行的病毒RNA分离将整合到试管实验室检测平台中。在探针设计和RNA富集之后，将在我们的封闭系统Liar分子分析仪原型中使用逆转录聚合酶链反应在不到1小时的时间内验证该系统的SCV检测灵敏度和SNP分析。我们相信这项拟议的研究将大大有助于NIAID的两个首要任务：1）提高我们国家的生物防御能力，2）加强传染病的诊断和治疗。

DESCRIPTION (provided by applicant): The goal of this research application is to develop a rapid, easy to use, integrated nucleic acid test for severe acute respiratory syndrome (SARS) based on our proprietary lab-in-a-tube (Liat TM) platform. The Liat system enables the integration of sample preparation, multiplex nucleic acid amplification and real-time quantification in a single closed tube system. The recent SARS epidemic showed that more sensitive, rapid, and automated nucleic acid testing capability is needed at the local level to better diagnose patients and improve quarantine effectiveness. When completed, the proposed system will be capable of taking raw plasma or sputum samples and performing fully automated sample-to-answer SARS associated coronavirus (SCV) nucleic acid testing at the point-of-care in less than 1 hour. In meeting these requirements, the system will have broad applications in bio-defense and civilian healthcare. To enable the Phase I goals several SCV-specific minor groove binding (MGB)-TaqMan probes will be designed and validated. Viral RNA isolation using silica magneti6 beads or nucleic acid hybridization methodologies will be integrated into the lab-in-a-tube testing platform. Following probe design and RNA enrichment, the system will be validated for SCV detection sensitivity and SNP analysis in less than 1 hour from sample-to-answer using reverse transcription-polymerase chain reaction in our closed system Liar Molecular Analyzer prototype. We believe this proposed research will contribute significantly to two of NIAID's top priorities: 1) improving our nation's bio-defense capability, and 2) enhancing the diagnosis and treatment of infectious diseases.