Engineering Research Center of Bio-process, MOE, School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, PR China.
Engineering Research Center of Bio-process, MOE, School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, PR China; College of Chemistry and Food Engineering, Changsha University of Science & Technology, Changsha 410114, China.
Food Chem. 2024 Mar 1;435:137587. doi: 10.1016/j.foodchem.2023.137587. Epub 2023 Sep 25.
Honey authenticity guarantee is crucial for consumer health and fair-trading commerce. New visual false-positive-free molecular lateral flow strip (LFS), termed 5'-3' exonuclease activity -directed false positive-eradicating PCR assisted lateral flow strip (FPE-PCR-LFS) was developed. This FPE-PCR-LFS explored the availability of using a signal-probe as the mediator to integrate the efficient amplification module with visual LFS module. With the genomic DNA extracted from target honey, the designed signal probe would be hydrolyzed and exhausted by the 5'-3' exonuclease activity of Taq DNA polymerase in the amplification process. The hydrolyzed signal probe would not be recognized and capture on the T line with only C line of LFS, reflecting the authenticity of the tested honey. And as low as 0.5% authenticity can be accurately identified in commercial honey samples. Significantly, the false-positive-interference was successfully eradicated for the final visual results judgement, which would greatly widen the application of molecular PCR-LFS in various fields.
蜂蜜真实性保证对消费者健康和公平贸易至关重要。新的视觉假阳性免费分子横向流动条（LFS），称为 5'-3'外切酶活性导向假阳性消除 PCR 辅助横向流动条（FPE-PCR-LFS）被开发出来。这种 FPE-PCR-LFS 探索了使用信号探针作为媒介将高效扩增模块与视觉 LFS 模块集成的可行性。用目标蜂蜜中提取的基因组 DNA，设计的信号探针将在扩增过程中被 Taq DNA 聚合酶的 5'-3'外切酶活性水解和耗尽。被水解的信号探针不会被识别和捕获在线上，只有 LFS 的线，反映了测试蜂蜜的真实性。并且在商业蜂蜜样本中可以准确识别低至 0.5%的真实性。显著的是，假阳性干扰被成功消除，用于最终的视觉结果判断，这将极大地拓宽分子 PCR-LFS 在各个领域的应用。
