| Objective:To develop a microfluidic chip chemiluminescence immunoassay?CLIA?system using droplet technology for rapid detection of infectious disease markers.Methods:We designed and fabricated a microfluidic chip chemiluminescence immunoassay system.The microfluidic chip consists of parallel tandem microwells connected with microslits.Due to hydrophobic surface of the microwells and the microslits,reagents needed for the immunoassay were preloaded in water-in-oil droplets in the microwells.Due to the surface tension effect,the microslits act as capillary valves to stop coalescence of adjacent droplets but allow the magnetic beads to pass through.Benefitted to this design,the multistep chemiluminescence immunoassay can be implemented in parallel through programmed magnetic manipulation.The microfluidic CLIA was applied for detection of two infectious disease markers,C-reactive protein?CRP?and Procalcitonin?PCT?.The results were compared with the Siemens instrument for methodological consistency analysis.Results:The developed microfluidic system can be applied for quantitative detection of CRP and PCT.Under optimized experimental conditions,all analytical procedures can be completed within 15 minutes.For the CRP assay,the linear detection range?LDR?was 0.8-200 ngˇmL-1?R2=0.993?with a limit of detection?LOD?of 0.31ngˇmL-1,and for the PCT assay,the LDR and LOD were 0.5-60 ngˇmL-1?R2=0.993?and 0.09 ngˇmL-1.The comparison test analysis indicates a high consistency between the microfluidic chip and the Siemens specific protein instrument.Conclusion:We develop a microfluidic chip CLIA system for detection of multiple infectious disease markers.This microfluidic system can simultaneously accomplish the quantitative detection assay of CRP and PCT with a detection precision close to the conventional methodology.The developed microfluidic CLIA system is well suited to Point-of-Care testing with advantages of easy control and rapid analysis. |
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