[Objective] In order to better understand the role of filamentous fungi in Chinese liquor fermentation process, a rapid and accurate method to monitor the change of fungal biomass is necessary. This study established a real-time qPCR method to detect and quantify Aspergillus tubingensis, which is widely used in Chinese liquor fermentation. [Methods] The methods of extracting genome from fermented grains have been optimized, the specific primers for A. tubingensis has also been designed and validated. [Results] The total DNA extracting from fermented grains can reach 1.060×105 ng/g by in situ mechanical crushing method. The applicability of real-time qPCR method to detect A. tubingensis in solid-state matrix has been validated in liquor fermentation process. [Conclusion] Real-time qPCR method can rapidly and accurately detect the fungal biomass in solid-state matrix, which provides a powerful tool for related researches.