Accuracy and stability enhanced honey authenticity visual tracing method via false positive-eradicating PCR assisted nucleic acid-capturing lateral flow strip

Abstract

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.