Establishment of a Real-time PCR Assay for Detection of Mold Fungi in Tobacco
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Abstract:
To establish a rapid and effective detection method for mold fungi in tobacco, a real-time PCR assay based on SYBR Green I was developed with a pair of primers designed according to the rDNA ITS1 gene of Aspergillus oryzae isolated from commercial tobacco. A recombinant T-vector containing the ITS1 gene was constructed as a positive control. The correlation coefficient of the standard curve was 0.998 and was highly sensitive with a detection limit of 101 copy/μL of positive recombinant plasmid. The technology was specific toward the genome of Aspergillus oryzae without any cross-reaction with other microbe genome. The developed real-time PCR method in this study can be used for detection of mold fungi in tobacco.
