Dual-mode sensor based on the synergy of magnetic separation and functionalized probes for the ultrasensitive detection of Clostridium perfringens

Abstract

Clostridium perfringens is an important foodborne pathogen, which has caused serious public health problems worldwide. So, there is an urgent need for rapid and ultrasensitive detection of C. perfringens. In this paper, a dual-mode sensing platform using the synergy between fluorescent and electrochemical signals for Clostridium perfringens detection was proposed. An electrochemical aptasensor was constructed by a dual-amplification technology based on a DNA walker and hybridization chain reaction (HCR). When the C. perfringens genomic DNA was present, it specifically bonded with FAM-labeled aptamer which triggered the DNA walker on hairpin DNA (hDNA) tracks to start the synthesis of double-stranded DNA. HCR occurred subsequently and produced long-chain DNA to absorb more methylene blue (MB). In this cycle, the fluorescent signals of released FAM-labeled aptamer could also be detected. The synergistic effects of MB and FAM significantly improved the sensitivity and accuracy of the dual-mode sensor. As a result, the biosensor displayed an excellent analytical performance for C. perfringens at a concentration of 1 to 10⁸ CFU g⁻¹. A minimum concentration of 1 CFU g⁻¹ and good accuracy were detected in real samples. The proposed ultrasensitive detection method for detecting C. perfringens in food showed great potential in controlling foodborne diseases.