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Infrared Plasmonic Biosensor with Tetrahedral DNA Nanostructure as Carriers for Label-Free and Ultrasensitive Detection of miR-155.

ABSTRACT: MicroRNAs play an important role in early development, cell proliferation, apoptosis, and cell death, and are aberrantly expressed in many types of cancers. To understand their function and diagnose cancer at an early stage, it is crucial to quantitatively detect microRNA without invasive labels. Here, a plasmonic biosensor based on surface-enhanced infrared absorption (SEIRA) for rapid, label-free, and ultrasensitive detection of miR-155 is reported. This technology leverages metamaterial perfect absorbers stimulating the SEIRA effect to provide up to 1000-fold near-field intensity enhancement over the microRNA fingerprint spectral bands. Additionally, it is discovered that the limit of detection (LOD) of the biosensor can be greatly improved by using tetrahedral DNA nanostructure (TDN) as carriers. By using near-field enhancement of SEIRA and specific binding of TDN, the biosensor achieves label-free detection of miR-155 with a high sensitivity of 1.162% pm^-1 and an excellent LOD of 100 × 10^-15 m. The LOD is about 5000 times lower than that using DNA single strand as probes and about 100 times lower than that of the fluorescence detection method. This work can not only provide a powerful diagnosis tool for the microRNAs detection but also gain new insights into the field of label-free and ultrasensitive SEIRA-based biosensing.

SUBMITTER: Hui X

PROVIDER: S-EPMC8373097 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BACKGROUND:DNA methylation detection is indispensable for the diagnosis and prognosis of various diseases including malignancies. Hence, it is crucial to develop a simple, sensitive, and specific detection strategy. METHODS:A novel fluorescent biosensor was developed based on a simple dual signal amplification strategy using functional dendritic DNA nanostructure and signal-enriching polystyrene microbeads in combination with ligase detection reaction (LDR). Dendritic DNA self-assembled from Y-DNA and X-DNA through enzyme-free DNA catalysis of a hairpin structure, which was prevented from unwinding at high temperature by adding psoralen. Then dendritic DNA polymer labeled with fluorescent dye Cy5 was ligated with reporter probe into a conjugate. Avidin-labeled polystyrene microbeads were specifically bound to biotin-labeled capture probe, and hybridized with target sequence and dendritic DNA. LDR was triggered by adding Taq ligase. When methylated cytosine existed, the capture probe and reporter probe labeled with fluorescent dye perfectly matched the target sequence, forming a stable duplex to generate a fluorescence signal. However, after bisulfite treatment, unmethylated cytosine was converted into uracil, resulting in a single base mismatch. No fluorescence signal was detected due to the absence of duplex. RESULTS:The obtained dendritic DNA polymer had a large volume. This method was time-saving and low-cost. Under the optimal experimental conditions using avidin-labeled polystyrene microbeads, the fluorescence signal was amplified more obviously, and DNA methylation was quantified ultrasensitively and selectively. The detection range of this sensor was 10-15 to 10-7 M, and the limit of detection reached as low as 0.4 fM. The constructed biosensor was also successfully used to analyze actual samples. CONCLUSION:This strategy has ultrasensitivity and high specificity for DNA methylation quantification, without requiring complex processes such as PCR and enzymatic digestion, which is thus of great value in tumor diagnosis and biomedical research.

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Infrared Plasmonic Biosensor with Tetrahedral DNA Nanostructure as Carriers for Label-Free and Ultrasensitive Detection of miR-155.

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