The invention discloses a multiple PCR detecting method for
pathogen of sugarcane mosaic
disease and ratoon stunting
disease, which is characterized in that (1), a sheathing leaf at the stem base portion of a sugarcane
plant is employed as detecting sample. (2), reverse transcription primer and primer pair of PCR detection cpc1 and cpc2 and RSD primer pair F1 and F2 are designed, sugarcane
mosaic virus and genetic fragment of
pathogen of sugarcane ratoon stunting
disease are amplified. (3), total
nucleic acid is extracted, and the
RNA thereof is turned into reverse transcription via cDNA. (4),
reaction system and amplification program of the one-
step method with multiple RT-PCR are designed, and the
nucleic acid is amplified. (5),
gel electrophoresis analysis and amplification results and specificity are employed, and simultaneously decuple progressive
dilution is used to detect the sensibility of the one-
step method with multiple RT-PCR. The detecting method provided by the invention accomplishes reverse transcription and PCR reaction and simultaneously detects
pathogen of two diseases by extracting the total
nucleic acid of sugarcane, which has the advantages of simple operation, economy of time, low cost, high specificity and sensibility and the like.