Design of a melting curve analysis (MCA) based on multiplex real-time PCR for detection of Aspergillus terreus and Aspergillus fumigatus in cereals and oilseeds samples

The positive control data analysis indicates that the lovB gene displays melt peaks between 86.2 °C and 86.8 °C. The easL gene shows peaks from 89.2 °C to 89.7 °C. A high cycle threshold was observed in 100 ng of DNA using 5-fold DNA dilutions of A. fumigatus NCPF 7097 and A. terreus ATCC 20,542. Also, 36 A. fumigatus isolates (25.7%) and 21 A. terreus isolates (15%) were recognized among the 140 cereals and oilseeds samples through phenotypic analysis. Furthermore, the MRT-PCR technique detected 24 isolates (17.1%) of A. fumigatus and 11 (7.8%) of A. terreus. The detection of Aspergillus species in cereals and oilseed samples can be successfully achieved by utilizing DNA from pure culture with MRT-RCR assay and MCA.