Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
866730 | Biosensors and Bioelectronics | 2014 | 5 Pages |
•Sensitive naked eye detection of cancer biomarkers is realized through a hybrid signal amplification approach.•Biobarcode amplification is coupled with enzyme-assisted DNA recycling to achieve hybrid signal enhancement.•Sub-picomolar carcinoembryonic antigen can be visually detected with the developed method.•The proposed method can be applied for visual detection of carcinoembryonic antigen in serum samples.
Naked eye-based detection has received increasing research interest due to the simplicity nature of this type of assay. However, improving the sensitivity of the naked eye detection method for the monitoring of trace amount of target molecules remains a major challenge. Herein, we describe a biobarcode and an enzyme-assisted DNA recycling hybrid amplification strategy for naked eye detection of sub-picomolar carcinoembryonic antigen (CEA), a cancer biomarker. The presence of CEA and the corresponding antibodies results in the formation of immunocomplexes and the capture of the biobarcodes in a microplate. The massive barcode DNAs released from the biobarcodes hybridize with the G-quadruplex inactive hairpin DNA probes and form catalytic nicking sites for N.BstNBI endonuclease, which cleaves the barcode DNA/hairpin partial dsDNA, releases the G-quadruplex active sequences and recycles the barcode DNA. Due to the barcode DNA recycling process, numerous G-quadruplex active sequences are generated and associate with hemin to form peroxidase mimicking enzymes, which convert colorless ABTS2− to green color intensified ABTS•− to achieve naked eye detection of CEA down to 0.025 ng mL−1 (0.14 pM). The naked eye detection strategy reported herein can be applied also to complicated serum sample matrix, making this approach hold great promise for point-of-care diagnostic applications.