Journal
BIOSENSORS & BIOELECTRONICS
Volume 92, Issue -, Pages 502-508Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2016.10.072
Keywords
RNAse H; Gold nanoparticles; Aggregation; DNA detection; Endonuclease enzyme
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Funding
- Department of Employment and Learning for Northern Ireland (DEL)
- Central Research Support Funds of Queen's University Belfast
- Agri-Food Quest Competence Centre R & D programme - Invest Northern Ireland Agency (Invest NI) [R3814GFS/RD1014267]
- Queen's University of Belfast AMR Network (QUBAN) - UK's Engineering and Physical Sciences Research Council (EPSRC) [EP/M027473/1]
- EPSRC [EP/M027473/1] Funding Source: UKRI
- Engineering and Physical Sciences Research Council [EP/M027473/1] Funding Source: researchfish
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The development of an ultrasensitive biosensor for the low-cost and on-site detection of pathogenic DNA could transform detection capabilities within food safety, environmental monitoring and clinical diagnosis. Herein, we present an innovative approach exploiting endonuclease-controlled aggregation of plasmonic gold nanoparticles (AuNPs) for label-free and ultrasensitive detection of bacterial DNA. The method utilizes RNA-functionalized AuNPs which form DNA-RNA heteroduplex structures through specific hybridization with target DNA. Once formed, the DNA-RNA heteroduplex is susceptible to RNAse H enzymatic cleavage of the RNA probe, allowing the target DNA to liberate and hybridize with another RNA probe. This continuously happens until all of the RNA probes are cleaved, leaving the nanoparticles unprotected and thus aggregated upon exposure to a high electrolytic medium. The assay is ultrasensitive, allowing the detection of target DNA at femtomolar level by simple spectroscopic analysis (40.7 fM and 2.45 fM as measured by UV-vis and dynamic light scattering (DLS), respectively). The target DNA spiked food matrix (chicken meat) is also successfully detected at a concentration of 1.2 pM (by UV-vis) or 18.0 fM (by DLS). In addition to the ultra-high sensitivity, the total analysis time of the assay is less than 3 h, thus demonstrating its practicality for food analysis.
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