Structurally responsive oligonucleotide-based single-probe lateral-flow test for detection of miRNA-21 mimics

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• 作者：Kamalodin Kor ; Anthony P. F. Turner ; Kobra Zarei…
• 关键词：miRNA ; Molecular beacon (MB) ; Structurally responsive assay ; Lateral ; flow strip tests ; DNA ; LNA probe
• 刊名：Analytical and Bioanalytical Chemistry
• 出版年：2016
• 出版时间：February 2016
• 年：2016
• 卷：408
• 期：5
• 页码：1475-1485
• 全文大小：2,776 KB
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• 作者单位：Kamalodin Kor (1) (2)
  Anthony P. F. Turner (1)
  Kobra Zarei (2)
  Morteza Atabati (2)
  Valerio Beni (1)
  Wing Cheung Mak (1)
  
  1. Biosensors and Bioelectronics Centre, Department of Physics, Chemistry and Biology (IFM), Linköping University, 58183, Linköping, Sweden
  2. School of Chemistry, Damghan University, Damghan, 36716-41167, Iran
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Analytical Chemistry
  Food Science
  Inorganic Chemistry
  Physical Chemistry
  Monitoring, Environmental Analysis and Environmental Ecotoxicology
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1618-2650

文摘

A single-probe strip test for the rapid and sensitive detection of miRNA-21 mimics is reported herein. Highly specific structurally responsive bi-functional, thiol and biotin, DNA/LNA oligonucleotide probes (molecular beacons-MB) were designed and conjugated with gold nanoparticles (AuNPs) (i.e. biotin-MB-AuNPs). The proposed design had the ability to modulate the accessibility of the biotin group as a function of the presence of a miRNA target allowing the interaction of the boilable with the streptavidin test zone only in the presence of the miRNA-21 mimics. For quantitative evaluation, images of the strip tests were recorded using a flatbed scanner (Epson Perfection V370 Photo). The colour intensities of the test zones of the strip tests were analysed with the ImageJ software (Scion Corp., USA) and quantified as a function of pixel intensity. The response of the strip test was linear over the range 0.5 to 20 nM miRNA-21 (limit of detection of 115 pM) and showed good reproducibility (intra and inter CVs below 8 %); furthermore, the assay was shown to be highly selective, discriminating other interference miRNAs mimics (e.g. miRNA-221 and miRNA-205). Finally, the proposed strip test was used for detection of miRNA-21 mimics in spiked serum samples, demonstrating its potential for point-of-care clinical applications. Main advantages of the single-probe strip test design are its versatility, simplicity and robustness, which can be easily extended to other miRNA targets by tuning the sequence of the single probe. Furthermore, the use of the structurally responsive single probe is particularly relevant in the case of short-length targets, such as miRNA, whereas a conventional sandwich approach might require a careful control of assay conditions such as hybridization temperature and salt concentration.