bDNA Microfluidic Disc for Accurate Monitoring of HIV-1

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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.