To address the problems of weak search ability, easily falling into local optimal solutions and poor path quality of sparrow search algorithm in AGV path planning, a multi-strategy improved sparrow search algorithm (MISSA) is proposed in this paper. MISSA improves the global search ability by improving the discoverer position update operator and introducing the sine cosine algorithm; adopts the adaptive number of vigilantes and adaptive adjustment step size to improve the convergence speed; introduces the Levy flight variation strategy to reduce the probability of falling into any local optimal solution; optimizes the boundary handling mechanism to prevent the loss of population diversity at a later stage; finally, uses the large-scale neighborhood search strategy and path smoothing mechanism for path optimization to further improve the path quality. The superiority-seeking ability of MISSA was verified by 12 standard test functions, and then 30 simulation experiments were conducted in grid maps with two specifications of 20×20 and 30×30. The experimental results showed that, by using MISSA, the path length was reduced by 44.1% and 63.1%, the number of turns was reduced by 68.4% and 78.4%, and the risk degree was reduced by 61.3% and 77.2%, which verifies the superiority of MISSA in path planning. Finally, MISSA was ported to the QBot2e mobile robot for physical verification to prove its feasibility in practical applications.