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機械式定量玻璃微粒免疫分析平台開發
Immunoassay detection is a technique for identifying diseases by measuring the concentration of specific antigent in a sample. It provides a simple and highly sensitive disease screening and treatment tracking for today's clinical medicine. The integration of immunoassay detection and microfluidic chip has become a very developmental technology, which can not only improve the shortcomings of large sample demand in traditional immunological methods, but also greatly shorten the detection time.
Among a variety of microfluidic immunoassay chips, the digital microfluidic chip has the advantages of less sample demand, faster detection speed, and high sensitivity. It uses the characteristics of a larger surface area per unit volume of particles to make it can react with the sample more quickly and evenly. At present, the mechanical glass particle digital microfluidic immunoassay chip developed by our laboratory uses a movable chip to improve the common defects of biological adhesion and low chip reliability in immunoassays. However, the analysis results have large variability and the measurement data stability still needs to be improved. Especially in the detection of a small amount of particles, the particle number has a great influence on the data, so this paper aims to develop a certain amount of particle immunoassay platform to improve the measurement accuracy of the particle-based digital microfluidic immunoassay chip.
In this study, the filter and particle quantitative chip can successfully fix the particle size range (40~60 um) and the initial particle number (25) in immunoassay, and combine with the mechanical glass particle digital microfluidic immunoassay chip for quantitative particle immunoassay. . Using human IL-1β as the assay antigen, the required sample size is 2 uL, the detection time is 42 minutes, the lowest detection limit (LOD) is 0.649 pg/mL, and the detection variance (CV value) of each concentration is all Less than 10%, the average detection variation is 4.78%. Compared the detection result obtained when the particles are not quantified (CV value 13%, LOD = 1.163 pg/mL), the particle quantitative immunoassay can reduce the minimum detection limit and the detection variation.