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  • Title: Ultrasensitive Label-Free Nucleic-Acid Biosensors Based on Bimodal Waveguide Interferometers.
    Author: Huertas CS, Lechuga LM.
    Journal: Methods Mol Biol; 2022; 2393():89-125. PubMed ID: 34837176.
    Abstract:
    The bimodal waveguide (BiMW) biosensor is an innovative common path interferometric sensor based on the evanescent field detection principle. This biosensor allows for the direct detection of virtually any biomolecular interaction in a label-free scheme by using specific biorecognition elements. Due to its inherent ultrasensitivity, it has been employed for the monitoring of relevant nucleic-acid sequences such as mRNA transcripts or microRNAs present at the attomolar-femtomolar concentration level in human samples. The application of the BiMW biosensor to detect these nucleic acids can be a powerful analytical tool for diagnosis and prognosis of complex illnesses, such as cancer, where these biomarkers play a major role. The BiMW sensor is fabricated using standard silicon-based microelectronics technology, which allows its miniaturization and cost-effective production, meeting the requirements of portability and disposability for the development of point-of-care (PoC) sensing platforms.In this chapter, we describe the working principle of the BiMW biosensor as well as its application for the analysis of nucleic acids. Concretely, we show a detailed description of DNA functionalization procedures and the complete analysis of two different RNA biomarkers for cancer diagnosis: (1) the analysis of mRNA transcripts generated by alternative splicing of Fas gene, and (2) the detection of miRNA 181a from urine liquid biopsies, for the early diagnosis of bladder cancer. The biosensing detection is performed by a direct assay in real time, by monitoring the changes in the intensity pattern of the light propagating through the BiMW biosensor, due to the hybridization of the target with the specific DNA probe previously functionalized on the BiMW sensor surface.
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