Methods: In this study, seven widely reported genes (GAPDH, TUBA, ACTB, SOD1, TP, ND and PS) of Schistosoma japonicum were systematically validated by combining Solexa high-throughput sequence analysis with targeted qPCR experiments to identify the most suitable reference genes on the 18th day and the 23rd day post infection of Schistosoma japonicum, and the expression stability of these seven candidate genes was then comprehensively evaluated using four complementary algorithms—the ΔCT method and the GeNorm V3.5, BestKeeper, and NormFinder software applications. The gene discussed is ACTB; the disease is infection.