TaqMan MGB-based real-time fluorescent PCR method for rapid detection of Setophoma terrestris
To accurately and rapidly detect the quarantine fungus Setophoma terrestris,a species-specific real-time polymerase chain reaction assay was developed in this study.A TaqMan-MGB probe was designed and synthesized according to the difference of rDNA ITS sequence between S.terrestris type strain and its closely related species.The whole reaction process took only about 1 h.The minimum detection limit was up to 1 pg/μL,and the optimized reaction conditions resulted in a final primer concentration of 0.6 μmol/L and a final probe concentration of 0.6 μmol/L.The method was highly accurate,rapid and sensitive,not only suitable for the detection of pure culture,but also for the detection of S.terrestris in onion tissue samples,providing a new reliable technological method for agroforestry production and customs inspection and quarantine.
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维普
