Point Detection of Pathogens in Oral Samples via UPT

描述（由申请人提供）：我们正在开发一种便携式、信用卡大小的微流体芯片实验室，用于使用口腔液体快速检测多种细菌和病毒靶标。在本研究的初始阶段，我们已经评估了一些口腔流体收集器，并选择了一个;设计和评估了5种类型的微型PCR装置，并选择了一个;设计和测试了几种类型的阀门，并将与两个进展，并评估了各种移动流体通过芯片的手段，并侧重于两种方法。该检测系统采用上转换荧光技术（UPT）和现有的阅读器。我们最初的工作集中在HIV（一种RNA病毒）和B。cereus（一种DNA细菌）来建立原理证明。该技术涉及模块化组件，可以进行修改以检测各种细菌和病毒以及这些病原体的抗体。在这次更新中，我们将继续开发一种监测病毒抗原、核酸和抗HIV抗体的HIV即时检测设备。我们的细菌研究将扩展到S。化脓性链球菌是导致链球菌性咽喉炎的主要微生物，选择它是因为它的高流行率有利于临床评价。我们将在2年内开发和优化能够同时检测细菌和病毒目标的原型便携式设备，并预计在5年内为完全符合FDA标准的临床试验做好准备。本提案中提出的研究/开发计划将（1）继续设计、测试和优化微流控芯片和处理单元;（2）使用来自HIV+和HIV-女性（WIHS）的库存口腔和血清样本以及来自S.化脓菌+和-个体，以评估该器械的稳健性、特异性、再现性和检测限;（3）评估几种提高灵敏度、减少分析时间和开发第二代器械的方法;（4）进行测定验证并制定完整的产品开发计划，包括设计控制程序。值得注意的是，虽然该计划的重点是艾滋病毒和S。通过检测口腔样本中的化脓性链球菌，该平台可以很容易地转换为其他类型的样本（血液、鼻咽和口腔拭子、尿液）和其他病原体，包括禽流感H5 N1病毒等新出现的传染性病原体。

DESCRIPTION (provided by applicant): We are developing a portable, credit-card sized microfluidic lab-on-a-chip for rapid point-of-care detection of multiple bacterial and viral targets using oral fluids. In the initial phase of this study we have evaluated a number of oral fluid collectors, and selected one; designed and evaluated 5 types of miniature PCR devices, and selected one; designed and tested several types of valves, and will progress with two, and assessed a variety of means of moving fluids through the chip and focused on two methods. The detection system utilizes Up-converting phosphor Technology (UPT) and an existing reader. Our initial work focused on HIV (an RNA virus) and B. cereus (a DNA bacterium) to establish proof of principle. The technology involves modular components, and can be modified to detect a wide variety of bacteria and viruses, as well as antibodies to those pathogens. In this renewal, we will continue development of an HIV point-of-care device that monitors viral antigen, nucleic acid, and anti-HIV antibodies. Our bacterial studies will be extended to S. pyogenes, the major organism responsible for strep throat, selected since its high prevalence facilitates clinical evaluation. We will develop and optimize a prototype portable device capable of simultaneously detecting bacterial and viral targets within 2 years, and anticipate having a system ready for full FDA compliant clinical trials in 5 years. The research/development plan presented in this proposal will (1) continue to design, test, and optimize the microfluidic chip and processing unit; (2) carry out clinical studies with banked oral and serum samples from HIV+ and HIV- women (WIHS) and oral swabs from S. pyogenes + and - individuals to assess the robustness, specificity, reproducibility and limits of detection of this device; (3) assess several approaches to enhance sensitivity, decrease analysis time, and lead to a second generation device; and (4) perform assay verification and develop a complete product development plan including a design control program. It is important to note that while the plan focuses on HIV and S. pyogenes in oral samples, the platform can easily be converted to look at other types of samples (blood, nasopharyngeal and oral swabs, urine), and other pathogens including emerging infectious agents such as avian H5N1 virus.