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，信用卡大小的微流控芯片实验室，用于使用口腔液体快速检测多种细菌和病毒目标。在这项研究的初始阶段，我们评估了一些口腔液体收集器，并选择了一个；设计和评价5种微型PCR装置，并筛选出1种；设计和测试了几种类型的阀门，并将对其中两种进行改进，并评估了通过芯片移动流体的各种方法，并重点研究了两种方法。检测系统利用上转换荧光粉技术（UPT）和现有的读取器。我们最初的工作集中在HIV（一种RNA病毒）和蜡样芽孢杆菌（一种DNA细菌）上，以建立原理证明。该技术包含模块化组件，可以进行修改以检测各种各样的细菌和病毒，以及针对这些病原体的抗体。在这次更新中，我们将继续开发一种监测病毒抗原、核酸和抗艾滋病毒抗体的艾滋病毒即时护理设备。我们的细菌研究将扩展到化脓性链球菌，这是导致链球菌性咽喉炎的主要微生物，选择化脓性链球菌是因为它的高患病率便于临床评估。我们将在2年内开发并优化一种能够同时检测细菌和病毒目标的便携式原型设备，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)对HIV+和HIV-妇女（WIHS）的口腔和血清样本以及化脓性链球菌+和-个体的口腔拭子进行临床研究，以评估该装置的稳健性、特异性、可重复性和检测局限性；(3)评估几种方法，以提高灵敏度，减少分析时间，并导致第二代设备；(4)进行分析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，虽然该计划侧重于口腔样本中的艾滋病毒和化脓性链球菌，但该平台可以很容易地转换为查看其他类型的样本（血液、鼻咽和口腔拭子、尿液）和其他病原体，包括新出现的传染性病原体，如禽流感H5N1病毒。

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.