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Cited by 292Advanced biosensing and bioanalysis techniquesBiosensors and Analytical DetectionDNA and Biological Computing

Real-time detection of isothermal amplification reactions with thermostable catalytic hairpin assembly

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Jiang, Yu (Sherry), Li, Bingling, Milligan, John N., Bhadra, Sanchita, Ellington, Andrew D.·2013·Journal of the American Chemical Society
DOI 10.1021/ja4023978PMC PMC3724415
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Abstract

Catalytic hairpin assembly (CHA) is an enzyme-free amplification method that has previously proven useful in amplifying and transducing signals at the terminus of nucleic acid amplification reactions. Here, for the first time, we engineered CHA to be thermostable from 37 to 60 °C and in consequence have generalized its application to the real-time detection of isothermal amplification reactions. CHA circuits were designed and optimized for both high- and low-temperature rolling circle amplification (RCA) and strand displacement amplification (SDA). The resulting circuits not only increased the specificity of detection but also improved the sensitivity by as much as 25- to 10000-fold over comparable real-time detection methods. These methods have been condensed into a set of general rules for the design of thermostable CHA circuits with high signals and low noise.

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  • The d-DNA hairpins d-H1 and d-H2 retain high stability under simulated physiological conditions (50 mM KCl, 20 mM NaCl, 1 mM MgCl2, pH 7.6, 37 °C) yet rapidly assemble into complex d-H1/H2 in the presence of initiator d-OUT155.

    “Ultimately, we identified a pair of hairpins, d-H1 and d-H2 (Table S1), which retained high stability under simulated physiological conditions (i.e., 50 mM KCl, 20 mM NaCl, 1 mM MgCl2, pH 7.6, 37 °C), yet rapidly assembled into complex d-H1/H2 the presence of the initiator strand (d-OUT155).”

    From l-DNA-Based Catalytic Hairpin Assembly Circuit (2020) · cites this paper as [32]

    In the section labeled Abstract (no page given), the source describes catalytic assembly of H1LT and H2LT and reports little background leakage at 37 °C. It does not identify d-H1, d-H2, or d-OUT155, or report their behavior under the specified simulated physiological conditions; its 37 °C circuit instead used 140 mM NaCl and 5 mM KCl at pH 7.5. (LLM, conf=0.82)

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Cited by

Sorted by citations

Authors
Jiang, Y. (., Li, B., Milligan, J. N., Bhadra, S., & Ellington, A. D.
Affiliations
The University of Texas at Austin (US)
Corresponding author
Ellington, Andrew D.
ORCID
Venue
Journal of the American Chemical Society
Publisher
American Chemical Society (ACS)
Published
2013-05-09
Volume
135
Issue
20
Pages
7430-7433
ISSN
  • 0002-7863 (ISSN-L)
  • 1520-5126
DOI
10.1021/ja4023978
PMC
PMC3724415
OpenAlex
W2066081299
Cited by
292
References
1
Citations by year
2026: 15 · 2025: 20 · 2024: 16 · 2023: 15 · 2022: 20 · 2021: 27 · 2020: 32 · 2019: 32 · 2018: 26 · 2017: 22 · 2016: 19 · 2015: 29 · 2014: 15 · 2013: 3
FWCI
9.86
Type
journal article
Language
en
Open access
Locations
  • Journal of the American Chemical Society (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • Foundation for the National Institutes of Health
  • Bill and Melinda Gates Foundation
  • Defense Advanced Research Projects Agency
  • National Institute of General Medical Sciences (R01GM094933)
  • National Institute of Allergy and Infectious Diseases (R01AI092839)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced biosensing and bioanalysis techniques
  • Physical Sciences › Engineering › Biomedical Engineering › Biosensors and Analytical Detection
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Biological Computing
Keywords
catalytic hairpin assembly, isothermal amplification, real-time detection, rolling circle amplification, strand displacement amplification, enzyme-free amplification, nucleic acid detection, signal transduction, detection sensitivity, detection specificity
MeSH terms
Catalysis, DNA, Nucleic Acid Hybridization, Temperature, Time Factors

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