The highest concentration of seven antibiotics on the growth of tobacco bacterial wilt, the inhabiting fungus and competitive bacterial were chosen with plate culture count. The results showed that ampicillin capsule was 9.996×10-5 μg/mL, ofloxacin tablets 9.990×10-7 μg/mL, roxithromycin tablets 2.998×10-6 μg/mL, cefradine capsules 3.000×10-6 μg/mL, acetylspiramycin tablets 1.998×10-5μg/mL, clarithromycin dispersible tablets 2.999×10-5 μg/mL, and azithromycin capsule was 1.999×10-6 μg/mL. The selected medium of the optimal concentration ratio of seven antibiotics to detect tobacco bacterial wilt in the soil from different areas was used, and it was found that the number of tobacco bacterial wilt in soil samples in Xiangxi was larger than in Yongzhou, and relatively smaller in Chenzhou. There was a positive correlation between the disease index and the number of bacterial wilt, and the tobacco bacteria in adding antagonistic bacterial fertilizer soil were less than in common soil. The method can be used to detect tobacco bacterial wilt quickly and to predict the occurrence of tobacco wilt as well.