Two-color, ultra-sensitive fluorescent strategy for ochratoxin A detection based on hybridization chain reaction and DNA tweezers

Abstract A two-color fluorescent DNA tweezers was developed for ultrasensitive detection of Ochratoxin A (OTA) based on hairpin-locked aptamer and hybridization chain reaction (HCR) amplification strategy. OTA can bind with hairpin-locked aptamer and then trigger the HCR reaction to produce a long double-strand DNA. The side-chains of the long duplex can separately hybridize with the two locker sequences of DNA tweezer, causing the opening of DNA tweezer and the recovery of two-color fluorescent signals. It shows a good linear range from 0.02 to 0.8 ppb with limit of detection of 0.006 ppb for FAM and 0.014 ppb for Cy5, which is beyond the requirement of actual application. In addition, the two-color fluorescent strategy can greatly reduce the false positive rate. It shows excellent performance for detection of OTA in practical food sample.