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Modified beacon probe assisted dual signal amplification for visual detection of microRNA.

Authors
  • Sun, Xiuwei1
  • Ying, Na2
  • Ju, Chuanjing3
  • Li, Zhongyi4
  • Xu, Na4
  • Qu, Guijuan5
  • Liu, Wensen6
  • Wan, Jiayu7
  • 1 Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China; College of Animal Science and Technology, Jilin Agricultural University, Changchun, China. , (China)
  • 2 East China Sea Fisheries Research Institute, China Academy of Fishery Sciences, Shanghai, China. , (China)
  • 3 Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China; The General Hospital of FAW, Changchun, China; The Fourth Hospital of Jilin University, Changchun, China. , (China)
  • 4 Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China. , (China)
  • 5 College of Animal Science and Technology, Jilin Agricultural University, Changchun, China. , (China)
  • 6 Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China. Electronic address: [email protected] , (China)
  • 7 Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China. Electronic address: [email protected] , (China)
Type
Published Article
Journal
Analytical Biochemistry
Publisher
Elsevier
Publication Date
Apr 21, 2018
Volume
550
Pages
68–71
Identifiers
DOI: 10.1016/j.ab.2018.04.010
PMID: 29684322
Source
Medline
Keywords
License
Unknown

Abstract

In a recent study, we reported a novel assay for the detection of microRNA-21 based on duplex-specific nuclease (DSN)-assisted isothermal cleavage and hybridization chain reaction (HCR) dual signal amplification. The Fam modified double-stranded DNA products were generated after the HCR, another biotin modified probe was digested by DSN and released from the magnetic beads after the addition of the target miRNA. The released sequence was then combined with HCR products to form a double-tagging dsDNA, which can be recognized by the lateral flow strips. In this study, we introduced a 2-OMethyl-RNA modified beacon probe (2-OMe-MB) to make some improvements based on the previous study. Firstly, the substitution of modified probe combined on magnetic beads avoids the fussy washing steps for the separation of un-reacted probes. Furthermore, the modification of 2-OMe on the stem of the probe avoided the unnecessary cleavage by DSN, which greatly reduce the background signal. Compared to the previous work, these improvements save us a lot of steps but possess the comparable sensitivity and selectivity.

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