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<p>Matcha, a premium powdered tea derived from <i>Camellia sinensis</i>, is prone to adulteration with non-tea plant materials due to its high market value, posing a serious challenge for food authenticity and the rights of consumers. Conventional morphological methods are unable to detect powdered adulterants, and spectroscopic methods exhibit limitations in detecting low-level adulterants. Therefore, this study developed a novel real-time PCR assay targeting the internal transcribed spacer 2 (ITS2) region for the rapid and sensitive detection of potential adulterants in matcha, including <i>Morus alba</i>, <i>Dianthus chinensis</i>, and <i>Hordeum vulgare</i>. The method showed target-specific amplification under the tested conditions and achieved detection limits as low as 0.1% (w/w) in artificial adulterant samples, corresponding to 0.1&#xa0;ng DNA. In commercial matcha samples, all samples exhibited positive amplification curves using the primer and probe of <i>C. sinensis</i>, confirming the presence of matcha DNA in all samples. Positive results were detected in 10 commercial samples for <i>M. alba</i> and in six samples for <i>H. vulgare</i> among the total 40 samples. And no positive results were observed for <i>D. chinensis</i> using target-specific primer and probe. All positive amplifications were further verified by Sanger sequencing and Basic Local Alignment Search Tool (BLAST) analysis, with sequence similarity exceeding 98% to reference genes. Overall, this real-time PCR-based assay provided a rapid, sensitive, and reliable tool for the detection of matcha adulteration, providing an effective tool for food quality control and regulatory surveillance.</p>

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Development of a Real-Time PCR Assay for the Detection of Plant-Based Adulterants in Chinese Green Tea Matcha Powder

  • 期刊论文

  • 发表日期：

    2026年4月15日

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  • Kunpeng Zhu,
  • Yi Wang,
  • Sijun Liu,
  • Fei Lao,
  • Ailiang Chen,
  • Aidong Sun,
  • Hui Li

摘要

Matcha, a premium powdered tea derived from Camellia sinensis, is prone to adulteration with non-tea plant materials due to its high market value, posing a serious challenge for food authenticity and the rights of consumers. Conventional morphological methods are unable to detect powdered adulterants, and spectroscopic methods exhibit limitations in detecting low-level adulterants. Therefore, this study developed a novel real-time PCR assay targeting the internal transcribed spacer 2 (ITS2) region for the rapid and sensitive detection of potential adulterants in matcha, including Morus alba, Dianthus chinensis, and Hordeum vulgare. The method showed target-specific amplification under the tested conditions and achieved detection limits as low as 0.1% (w/w) in artificial adulterant samples, corresponding to 0.1 ng DNA. In commercial matcha samples, all samples exhibited positive amplification curves using the primer and probe of C. sinensis, confirming the presence of matcha DNA in all samples. Positive results were detected in 10 commercial samples for M. alba and in six samples for H. vulgare among the total 40 samples. And no positive results were observed for D. chinensis using target-specific primer and probe. All positive amplifications were further verified by Sanger sequencing and Basic Local Alignment Search Tool (BLAST) analysis, with sequence similarity exceeding 98% to reference genes. Overall, this real-time PCR-based assay provided a rapid, sensitive, and reliable tool for the detection of matcha adulteration, providing an effective tool for food quality control and regulatory surveillance.