Ochratoxin A (OTA) is a polyketide mycotoxin that is produced by Aspergillus and Penicillium. Food contaminated with OTA poses health risks and is a food-safety challenge. Quantitative polymerase chain reaction (qPCR) has been used to identify non-toxigenic and toxigenic strains from coffee samples using polyketide synthase (pks), the OTA synthesis gene. In this research, Aspergillus carbonarius (ochratoxin-producing strain) and A. flavus (non-ochratoxin-producing strain) were used to amplify a 141 bp fragment of the pks gene. The 141 bp PCR product was successfully cloned into TOPO ® TA plasmid. Subsequently, ten-fold dilutions of plasmid DNA were used to generate the standard curve by plotting the threshold cycle against log DNA concentration using qPCR. Further, fungal DNA contamination was quantified in 11 samples of roasted coffee using qPCR. All 11 coffee samples were accepted as safe, since the fungal genomic DNA contamination was less than 3.85 x 10 3 copies. Therefore, this research suggested that qPCR is a fast and accurate method to detect and quantify OTA-producing fungi in coffee products. Thus, we successfully developed a system to quantify fungal contamination in coffee.
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