【Aim】Alternaria triticina causing wheat leaf blight is a quarantine fungus in China. This study establishes aTaqMan-MGB probe-based real-time PCR method targeting species-specific genetic markers for rapid, sensitive, and accurate detection of this fungal pathogen. 【Method】 According to the sequence differences in the translation elongation factor (EF) betweenAlternaria triticina and its closely related species, a pair of species-specific primers and a minor groove binder (MGB) probe were designed via conserved region alignments. The real-time PCR conditions were optimized as follows: a final primer concentration of 0.7 μmol·L^-1 and a final probe concentration of 0.9 μmol·L^-1. The feasibility of the method was validated through specificity, sensitivity, and simulated sample testing. 【Result】 Specificity testing results demonstrated that the method can specifically detect A. triticina while showing no cross-reactivity with non-target species. Sensitivity testing results showed that the lowest detection limit was 20 pg of total DNA in a 10 μL reaction system. Simulated sample testing results confirmed the methods applicability for detecting samples suspected of carrying A. triticina. The entire reaction process took approximately 1 h, and the detection was performed in one-tube without post-PCR handling. 【Conclusion】 The optimized real-time PCR detection method developed in this study is rapid, sensitive, and accurate. It provides a crucial technical reference for the early detection of A. triticina, which is critical for preventing the introduction and spread of this major wheat pathogen, supporting enhanced quarantine surveillance programs.