摘要： Label-free, direct, rapid and real-time quantification method for human enterovirus 71 (EV71) using surface plasmon resonance (SPR) sensor is presented in this study. The performance of the four layer sensor chip was compared with the conventional Au monolayer sensor chip. Four layer sensor chip gave optimum resonance wavelength around 610 nm, which was same with the OLED light source peak wavelength and gave better quality factor (Q = DIP/FWHM). The result showed that the structure with 3 nm Al, 10 nm Au, 20 nm Ag, 10 nm Au layer sensor chip gave better limit of detection than the 3 nm Cr, and 47 nm Au layer sensor chip. The detection limit of direct quantification of EV71 particles is improved to 43 vp/mL of EV71 in DMEM medium. The results proved that the SPR biosensor with four layer sensor chip structure demonstrated great performance in the quantification of EV71 virus species.