An fluorescence resonance energy transfer sensing platform based on signal amplification strategy of hybridization chain reaction and triplex DNA for the detection of Chloramphenicol in milk

The use of chloramphenicol (CAP) in food had been strictly regulated or banned in many countries. Herein, an enzyme-free fluorescence resonance energy transfer (FRET) strategy was established for sensitive, rapid and specific detection of CAP in milk, which was based on triplex DNA and hybridization chain reaction amplification. CAP can specifically bind to the aptamer and release the trigger sequence, causing HCR to efficiently prime and forming triplex DNA, hence the FRET pairs (FAM and TAMRA) were close enough to cause fluorescent decreases. Consequently, CAP can be quantitatively detected by measuring the fluorescence reduction at 520 nm, and the reliability of the method was confirmed by enzyme-linked immunosorbent assay. The limit of CAP detection for 1.2 pg.mL- 1, and the average recoveries of milk samples were 97.5%-106%, and the relative standard deviation were 3.9%-5.3%. Thus, this method has a wide range of potential applications in CAP detection.

Automated summary

An enzyme-free fluorescence resonance energy transfer (FRET) strategy based on triplex DNA and hybridization chain reaction amplification was established for sensitive detection of chloramphenicol (CAP) in milk. This method allows for accurate and rapid quantification of CAP, with a wide range of potential applications.