In high-reliability applications, the health condition of the MEMS gyroscope needs to be known in real time to ensure that the system does not fail due to the wrong output signal. Because the MEMS gyroscope self-test based on the principle of electrostatic force cannot be performed during the working state. We propose that by monitoring the quadrature error signal of the MEMS gyroscope in real time, an online self-test of the MEMS gyroscope can be realized. The correlation between the gyroscope’s quadrature error amplitude signal and the gyroscope scale factor and bias was theoretically analyzed. Based on the sixteen-sided cobweb-like MEMS gyroscope, the real-time built-in self-test (BIST) method of the MEMS gyroscope based on the quadrature error signal was verified. By artificially setting the control signal of the gyroscope to zero, we imitated several scenarios where the gyroscope malfunctioned. Moreover, a mechanical impact table was used to impact the gyroscope. After a 6000 g shock, the gyroscope scale factor, bias, and quadrature error amplitude changed by −1.02%, −5.76%, and −3.74%, respectively, compared to before the impact. The gyroscope failed after a 10,000 g impact, and the quadrature error amplitude changed −99.82% compared to before the impact. The experimental results show that, when the amplitude of the quadrature error signal seriously deviates from the original value, it can be determined that the gyroscope output signal is invalid.