Point-of-Care Testing(POCT),a rapid diagnostic testing performed at the sites near patients,has recently aroused great interest for solving problems in global health.The research and innovation of microfluidic paper-based analytic device(μPAD)for POCT is particularly emphasized due to its advantages of portability,low-cost and disposability.Here,we describe a versatile qualitative POCT platform for simultaneous detection of multiple targets based on target-mediated "stop-flow" hydrogel assembled μPAD.Based on the specific combination between target and corresponding aptamer,two models from hydrogel-stop and solution-flow are regulated by targets-mediation with different final displacement.In the absence of target,sample solution will re-dissolve L-Apt and SA & SB filled in μPAD before to activate the hydrogel reaction with a stop-flow of the sample.Conversely,in the presence of target,a solution-flow model is obtained through the open-reaction of target and L-Apt without the formation of hydrogel.The testing result can be achieved in 6 min by naked eyes via a simple loading operation without any accessorial equipment.In this assay,we realize a rapid and multiplex qualitative detection for cocaine,adenosine and Pb2+ simultaneously.In addition,the approach enables to work in complex biological matrixes such as urine.In view of the rapid and user-friendly detection by our approach,a potential application prospect and commercial value is expected.