The invention discloses a combination of primers and probes for detection of carbendazim-resistant strains of Fusarium graminearum and a detection method thereof. The primer-probe combination consists of forward primer tub2F, reverse primer tub2R, three drug-resistant mutants and wild-type probes 167m, 198m, 200m, 167w, 198w, and 200w. The detection method of the present invention can simultaneously detect three different types of drug-resistant mutations, and has high throughput, strong specificity and scalability, high signal-to-noise ratio, high sensitivity, high degree of automation, strong practicability, and can meet the requirements of anti-carbendazim and The rapid detection of Fusarium valley provides a basis for scientific and rational drug use.