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<p>The increasing prevalence of food allergies worldwide underscores an urgent need for detection methods that are swift, accurate, and dependable. While traditional protein-based immunoassays and DNA-based PCR are widely used, they are hampered by limitations such as protein denaturation, cross-reactivity, and reliance on sophisticated thermal cycling equipment. This review comprehensively examines the field of isothermal nucleic acid amplification techniques (INAATs) as promising complementary tools for food allergen detection. We elucidate the fundamental principles of key INAAT platforms, encompassing both enzyme-dependent and enzyme-free methods that have found utility in this field. The article systematically synthesizes the application of these techniques for detecting all major food allergens (“Big Eight” and beyond), categorizing strategies into DNA-based detection of species-specific genes and INAAT-powered signal amplification in protein-based biosensors using aptamers or antibodies. Our analysis shows that INAATs offer notable strengths in sensitivity, specificity, rapidity, and operational simplicity, making them well-suited for on-site and point-of-need testing. We also objectively discuss persistent challenges, including susceptibility to false positives from aerosol contamination, difficulties in reliable quantification and inter-laboratory reproducibility, the scarcity of certified reference materials, reagent stability constraints for field deployment, and the absence of standardized protocols and regulatory validation frameworks. Finally, the review outlines future directions that are closely linked to these limitations: multiplexing, integration with CRISPR-Cas systems, development of fully automated “sample-in-answer-out” platforms, and crucially, the establishment of standardized guidelines and formal validation through multi‑center trials. While INAATs are not a universal replacement for established methods, they constitute a promising complementary strategy, provided that technical and standardization hurdles are adequately addressed. This review aims to offer a balanced, evidence-based resource for researchers and end-users navigating the evolving landscape of allergen diagnostics.</p>

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Advancing Food Safety: The Role of Isothermal Nucleic Acid Amplification Techniques in Allergen Detection

  • 期刊论文

  • 发表日期：

    2026年7月25日

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  • Yingying Chen,
  • Wei Wang,
  • Ruifeng Mao

摘要

The increasing prevalence of food allergies worldwide underscores an urgent need for detection methods that are swift, accurate, and dependable. While traditional protein-based immunoassays and DNA-based PCR are widely used, they are hampered by limitations such as protein denaturation, cross-reactivity, and reliance on sophisticated thermal cycling equipment. This review comprehensively examines the field of isothermal nucleic acid amplification techniques (INAATs) as promising complementary tools for food allergen detection. We elucidate the fundamental principles of key INAAT platforms, encompassing both enzyme-dependent and enzyme-free methods that have found utility in this field. The article systematically synthesizes the application of these techniques for detecting all major food allergens (“Big Eight” and beyond), categorizing strategies into DNA-based detection of species-specific genes and INAAT-powered signal amplification in protein-based biosensors using aptamers or antibodies. Our analysis shows that INAATs offer notable strengths in sensitivity, specificity, rapidity, and operational simplicity, making them well-suited for on-site and point-of-need testing. We also objectively discuss persistent challenges, including susceptibility to false positives from aerosol contamination, difficulties in reliable quantification and inter-laboratory reproducibility, the scarcity of certified reference materials, reagent stability constraints for field deployment, and the absence of standardized protocols and regulatory validation frameworks. Finally, the review outlines future directions that are closely linked to these limitations: multiplexing, integration with CRISPR-Cas systems, development of fully automated “sample-in-answer-out” platforms, and crucially, the establishment of standardized guidelines and formal validation through multi‑center trials. While INAATs are not a universal replacement for established methods, they constitute a promising complementary strategy, provided that technical and standardization hurdles are adequately addressed. This review aims to offer a balanced, evidence-based resource for researchers and end-users navigating the evolving landscape of allergen diagnostics.