This paper proposes a method of fault detection of the connection of Lithium-Ion batteries based on entropy for electric vehicle. In electric vehicle operation process, some factors, such as road conditions, driving habits, vehicle performance, always affect batteries by vibration, which easily cause loosing or virtual connection between batteries. Through the simulation of the battery charging and discharging experiment under vibration environment, the data of voltage fluctuation can be obtained. Meanwhile, an optimal filtering method is adopted using discrete cosine filter method to analyze the characteristics of system noise, based on the voltage set when batteries are working under different vibration frequency. Experimental data processed by filtering is analyzed based on local Shannon entropy, ensemble Shannon entropy and sample entropy. And the best way to find a method of fault detection of the connection of lithium-ion batteries based on entropy is presented for electric vehicle. The experimental data shows that ensemble Shannon entropy can predict the accurate time and the location of battery connection failure in real time. Besides electric-vehicle industry, this method can also be used in other areas in complex vibration environment.