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      Biosensing Amplification by Hybridization Chain Reaction on Phase-Sensitive Surface Plasmon Resonance

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          Abstract

          Surface Plasmon Resonance (SPR) is widely used in biological and chemical sensing with fascinating properties. However, the application of SPR to detect trace targets is hampered by non-specific binding and poor signal. A variety of approaches for amplification have been explored to overcome this deficiency including DNA aptamers as versatile target detection tools. Hybridization chain reaction (HCR) is a high-efficiency enzyme-free DNA amplification method operated at room temperature, in which two stable species of DNA hairpins coexist in solution until the introduction of the initiator strand triggers a cascade of hybridization events. At an optimal salt condition, as the concentrations of H1 and H2 increased, the HCR signals were enhanced, leading to signal amplification reaching up to 6.5-fold of the detection measure at 30 min. This feature enables DNA to act as an amplifying transducer for biosensing applications to provide an enzyme-free alternative that can easily detect complex DNA sequences. Improvement of more diverse recognition events can be achieved by integrating HCR with a phase-sensitive SPR (pSPR)-tested aptamer stimulus. This work seeks to establish pSPR aptamer system for highly informative sensing by means of an amplification HCR. Thus, combining pSPR and HCR technologies provide an expandable platform for sensitive biosensing.

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          Most cited references43

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          Tumour exosome integrins determine organotropic metastasis

          Ever since Stephen Paget’s 1889 hypothesis, metastatic organotropism has remained one of cancer’s greatest mysteries. Here we demonstrate that exosomes from mouse and human lung-, liver- and brain-tropic tumour cells fuse preferentially with resident cells at their predicted destination, namely lung fibroblasts and epithelial cells, liver Kupffer cells and brain endothelial cells. We show that tumour-derived exosomes uptaken by organ-specific cells prepare the pre-metastatic niche. Treatment with exosomes from lung-tropic models redirected the metastasis of bone-tropic tumour cells. Exosome proteomics revealed distinct integrin expression patterns, in which the exosomal integrins α6β4 and α6β1 were associated with lung metastasis, while exosomal integrin αvβ5 was linked to liver metastasis. Targeting the integrins α6β4 and αvβ5 decreased exosome uptake, as well as lung and liver metastasis, respectively. We demonstrate that exosome integrin uptake by resident cells activates Src phosphorylation and pro-inflammatory S100 gene expression. Finally, our clinical data indicate that exosomal integrins could be used to predict organ-specific metastasis.
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            Surface plasmon resonance sensors for detection of chemical and biological species.

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              Real-time quantification of microRNAs by stem–loop RT–PCR

              A novel microRNA (miRNA) quantification method has been developed using stem–loop RT followed by TaqMan PCR analysis. Stem–loop RT primers are better than conventional ones in terms of RT efficiency and specificity. TaqMan miRNA assays are specific for mature miRNAs and discriminate among related miRNAs that differ by as little as one nucleotide. Furthermore, they are not affected by genomic DNA contamination. Precise quantification is achieved routinely with as little as 25 pg of total RNA for most miRNAs. In fact, the high sensitivity, specificity and precision of this method allows for direct analysis of a single cell without nucleic acid purification. Like standard TaqMan gene expression assays, TaqMan miRNA assays exhibit a dynamic range of seven orders of magnitude. Quantification of five miRNAs in seven mouse tissues showed variation from less than 10 to more than 30 000 copies per cell. This method enables fast, accurate and sensitive miRNA expression profiling and can identify and monitor potential biomarkers specific to tissues or diseases. Stem–loop RT–PCR can be used for the quantification of other small RNA molecules such as short interfering RNAs (siRNAs). Furthermore, the concept of stem–loop RT primer design could be applied in small RNA cloning and multiplex assays for better specificity and efficiency.
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                Author and article information

                Journal
                Biosensors (Basel)
                Biosensors (Basel)
                biosensors
                Biosensors
                MDPI
                2079-6374
                06 March 2021
                March 2021
                : 11
                : 3
                : 75
                Affiliations
                [1 ]Graduate Institute of Bioelectronics and Bioinformatics, National Taiwan University, Taipei 106, Taiwan; d02945009@ 123456ntu.edu.tw (C.-H.Y.); aresation@ 123456gmail.com (T.-H.W.)
                [2 ]Department of Medical Biotechnology and Laboratory Sciences, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan; chang@ 123456mail.cgu.edu.tw
                [3 ]Kidney Research Center, Department of Nephrology, Chang Gung Memorial Hospital, Taoyuan 333, Taiwan
                [4 ]Department of Biomedical Engineering, National Taiwan University, Taipei 106, Taiwan; onlyby1231@ 123456gmail.com (H.-Y.L.); k8006780067@ 123456yahoo.com.tw (H.-W.L.)
                [5 ]Department of Electrical Engineering, National Taiwan University, Taipei 106, Taiwan
                [6 ]Biomedical Technology and Device Research Laboratories, Industrial Technology Research Institute, Hsinchu 310, Taiwan
                Author notes
                [* ]Correspondence: nthuang@ 123456ntu.edu.tw (N.-T.H.); cwlinx@ 123456ntu.edu.tw (C.-W.L.)
                Author information
                https://orcid.org/0000-0001-5466-4724
                https://orcid.org/0000-0002-2569-805X
                Article
                biosensors-11-00075
                10.3390/bios11030075
                7998988
                5c2b79f5-f186-428a-a064-863be8fe344c
                © 2021 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 11 February 2021
                : 02 March 2021
                Categories
                Article

                aptamer,hybridization chain reaction,phase-sensitive surface plasmon resonance (pspr) biosensor

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