bDNA Microfluidic Disc for Accurate Monitoring of HIV-1

描述（由申请人提供）：需要更稳健的HIV-1定量检测方法，以便更好地预测患者预后和准确监测抗病毒治疗。分支DNA（bDNA）技术是一种线性信号放大方法，是检测中高拷贝数的最具重现性的方法（P < 0.001）。但目前针对低HIV RNA拷贝数（< 500拷贝/ml）的bDNA检测仍然缺乏准确性（CV = 37%），无法可靠地用于临床环境中的短期和长期病毒载量监测。更重要的是，我们人群中44%的标本显示HIV-1 RNA水平低于400拷贝/ml。该项目提出开发一种独立的微流控bDNA盘（mbDNA-Disc），具有非常可靠和自动化的微流控机制，用于快速，准确和灵敏的低HIV-1 RNA定量。基于双重扩增原理、bDNA多聚体和酶化学发光，mbDNA系统利用微流体技术提供所有必要的核酸化学反应，因为微流体技术已被证明可以提高许多生化过程的重现性和成本效益。在第一阶段项目中，我们将设计mbDNA-Disc配置，构建HIV-微流控盘和微致动器，集成和表征mbDNA-Disc系统，测试样品，并评估技术可行性。

DESCRIPTION (provided by applicant): A need exists for more robust assays for the quantification of HIV-1 for better prognosis of patients and accurate monitoring of antiviral therapy. Branched DMA (bDNA) technology, a linear signal amplification method, is the most reproducible method for detection of medium to high copy numbers (P < 0.001). But current bDNA assays for low HIV RNA copy numbers (< 500 copies/ml) still lack accuracy (CV = 37%) to be reliably used in a clinical setting for short- and long-term viral load monitoring. More importantly, 44% of specimens in our population show HIV-1 RNA levels below 400 copies/ml. This project proposes to develop a self-contained microfluidic bDNA disc (mbDNA-Disc) with a very reliable and automated microfluidic mechanism for rapid, accurate, and sensitive low HIV-1 RNA quantification. Based on the double amplification principle, bDNA multimer, and enzyme chemiluminescence, the mbDNA system utilizes microfluidics to provide all necessary nucleic acid chemistries, as microfluidics has proven to increase reproducibility and cost-efficiency for numerous biochemical processes. During the Phase I project, we will design the mbDNA-Disc configuration, construct the HIV-microfluidic disc and microactuator, integrate and characterize the mbDNA-Disc system, test sample, and evaluate technical feasibility.