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<p>This study aimed to develop a real-time PCR assay to specifically distinguish between <i>L. angustifolius</i> and <i>L. albus</i>. Species-specific primers were designed based on sequence variations in chloroplast DNA, including the <i>rps16-accD</i>, <i>matK, rbcL</i>, and <i>accD</i> regions, to target DNA from two species using six primers. The primers demonstrated high specificity, successfully differentiating the two species without cross-reactivity. The assay exhibited strong linearity and consistent amplification across a wide range, with a detection limit as low as 0.1&#xa0;pg. No false-positive or false-negative results were observed. Application to commercially available food samples confirmed the practical utility of the assay for detecting lupin DNA in distributed products. These findings indicate that the developed assay is a robust and efficient tool for species authentication, contributing to improved traceability, allergen management, and regulatory compliance in lupin-containing food systems.</p>

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Development of species-specific SYBR green-based real-time PCR assay to ifferentiate between Lupinus angustifolius and Lupinus albus

  • 期刊论文

  • 发表日期：

    2026年6月16日

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  • Ayoung Cho,
  • Ji Seok Yi,
  • Yun Seo Jung,
  • Ho Soo Lim,
  • Moon Ik Chang

摘要

This study aimed to develop a real-time PCR assay to specifically distinguish between L. angustifolius and L. albus. Species-specific primers were designed based on sequence variations in chloroplast DNA, including the rps16-accD, matK, rbcL, and accD regions, to target DNA from two species using six primers. The primers demonstrated high specificity, successfully differentiating the two species without cross-reactivity. The assay exhibited strong linearity and consistent amplification across a wide range, with a detection limit as low as 0.1 pg. No false-positive or false-negative results were observed. Application to commercially available food samples confirmed the practical utility of the assay for detecting lupin DNA in distributed products. These findings indicate that the developed assay is a robust and efficient tool for species authentication, contributing to improved traceability, allergen management, and regulatory compliance in lupin-containing food systems.