Abstract: 【Objective】Hami melon bacterial fruit blotch (HBFB) is a new disease in China, the pathogen is Acidovorax avenae subsp. citrulli, which damages the fruits and lowers their qualities. HBFB is a typically seedborne bacterial disease, and the important prevention and cure method is the quarantine of the seeds with bacteria.【Method】and【Result】In this study, the immuno-detection was combined with PCR by using the superparamagnetic beads to form IMS-Realtime-PCR, which increase the precision of bacteria detection to app. 3×102 CFU/ml. The integrated method of bacterial detection was formed through combining the ASCM culturing, IMS and real-time PCR. 【Conclusion】One seed with bacteria in 1000 seeds could be detected using this integrated method.