The invention discloses a miRNA indirect real-time fluorescence quantitative PCR detection method. The method includes: firstly using single stranded DNA and miRNA for combination, then using DSN enzyme for digestion of paired single stranded DNA, and finally performing quantification on the remaining single stranded DNA by Taqman method, thus indirectly achieving detection of the target miRNA content. The method provided by the invention avoids the non-specific amplification problem in amplification of the traditional method, still uses the mature Taqman technology, and is very convenient and stable in application. Based on the above advantages, the method has a wide application prospect in the field of tumor detection.