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Objective <p>In the early stages of food production, detection of Aspergillus species contamination is crucial to prevent potential harm to human health. The primary focus of this study was to evaluate the performance of Multiplex Real-Time PCR (MRT-PCR) and melting curve analysis (MCA) in identifying <i>Aspergillus terreus</i> and <i>Aspergillus fumigatus</i> in cereals and oilseeds.</p> Results <p>The positive control data analysis indicates that the <i>lovB</i> gene displays melt peaks between 86.2&#xa0;°C and 86.8&#xa0;°C. The <i>easL</i> gene shows peaks from 89.2&#xa0;°C to 89.7&#xa0;°C. A high cycle threshold was observed in 100 ng of DNA using 5-fold DNA dilutions of <i>A. fumigatus</i> NCPF 7097 and <i>A. terreus</i> ATCC 20,542. Also, 36 <i>A. fumigatus</i> isolates (25.7%) and 21 <i>A. terreus</i> 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 <i>A. fumigatus</i> and 11 (7.8%) of <i>A. terreus</i>. 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.</p>

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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

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年7月14日

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  • Mahtab Garousin,
  • Reza Habibipour

摘要

Objective

In the early stages of food production, detection of Aspergillus species contamination is crucial to prevent potential harm to human health. The primary focus of this study was to evaluate the performance of Multiplex Real-Time PCR (MRT-PCR) and melting curve analysis (MCA) in identifying Aspergillus terreus and Aspergillus fumigatus in cereals and oilseeds.

Results

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.