We present a learning-based Shack-Hartmann wavefront sensor (SHWS) to achieve the high-order aberration detection without image segmentation or centroid positioning. Zernike coefficient amplitudes of aberrations measured from biological samples are referred and expanded to generate the training datasets. With one SHWS pattern inputted, up to 120th Zernike modes could be predicted within 10.9 ms with 95.56% model accuracy by a personal computer. The statistical experimental results show that compared with traditional modal-based SHWS, the root mean squared error in phase residuals of this method is reduced by ∼40.54% and the Strehl ratio of the point spread functions is improved by ∼27.31%. The aberration detection performance of this method is also validated on a mouse brain slice with 300 µm thickness and the median improvement of peak-to-background ratio of this method is ∼30% to 40% compared with traditional SHWS. With the high detection accuracy, simple processes, fast prediction speed and good compatibility, this work offers a potential approach to improve the wavefront sensing ability of SHWS, which could be combined with an existing adaptive optics system and be further applied in biological applications.