A power-free sample-loading microfluidic chip integrated with a colorimetric loop-mediated isothermal amplification and a chromatic analysis via image processing for multiplex detection of parasitic diseases.

The gold standard diagnostic methods of gastrointestinal helminth parasites are the identification based on morphological characters of adult worms and the separation of the egg stage from host feces using the flotation or sedimentation techniques, both of which are time-consuming and require technical skills and parasitic knowledge. In our study, we develop a power-free sample-loading microfluidic chip integrated with a colorimetric loop-mediated isothermal amplification (LAMP) and a chromatic analysis for multiplex detection of parasitic diseases. The microfluidic chip can detect five parasitic groups simultaneously. The results can be interpreted with the naked eye or by chromatic analysis via image processing. The analytical specificity of this assay is specific to R. echinobothrida, R. tetragona, R. cesticillus, A. galli, and the Echinostomatidae group, and had minimum DNA detectable concentrations ranging from 4 ng to 40 pg/chip. The performance of the developed assay on fecal samples revealed a strong agreement with the gold standard method (Cohen's kappa value = 0.9); McNemar's test indicated no significant differences between both assays (p ‍> ‍0.05). Consequently, the proposed assay has potential to apply the detection of a wide range of pathogens and infectious diseases.