ArticleBiochemistry2008
Specific versus nonspecific isothermal DNA amplification through thermophilic polymerase and nicking enzyme activities.
Article in Biochemistry, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 40 papers.
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Who cites it
40 citing papers in PubMed.
- Assessment of salivary microRNA by RT-qPCR: Facing challenges in data interpretation for clinical diagnosis.PloS one · 2024Trial
- Coupling Exponential Rolling Circle Amplification to Lateral Flow Devices for Point-of-Care Detection of Viral RNA.Angewandte Chemie (International ed. in English) · 2026Article
- One-Pot Detection of miRNA by Dual Rolling Circle Amplification at Ambient Temperature with High Specificity and Sensitivity.Biosensors · 2025Article
- Robust Sequence Design Space for the Isothermal Exponential Amplification of Short Oligonucleotides.Small (Weinheim an der Bergstrasse, Germany) · 2024Article
- Single G-quadruplex-based fluorescence method for the uracil-DNA glycosylase inhibitor screening.Heliyon · 2024Article
- Isothermal exponential amplification reactions triggered by circular templates (cEXPAR) targeting miRNA.Molecular biology reports · 2023Article
- Suitability evaluation of toehold switch and EXPAR for cell-free MicroRNA biosensor development.Biotechnology notes (Amsterdam, Netherlands) · 2023Article
- Cell-Free Production Systems in Droplet Microfluidics.Advances in biochemical engineering/biotechnology · 2023Review
- Positive feedback drives a secondary nonlinear product burst during a biphasic DNA amplification reaction.The Analyst · 2022Article
- A small-molecule chemical interface for molecular programs.Nucleic acids research · 2021Article
- Three-Way Junction-Induced Isothermal Amplification with High Signal-to-Background Ratio for Detection of Pathogenic Bacteria.Sensors (Basel, Switzerland) · 2021Article
- Developments in integrating nucleic acid isothermal amplification and detection systems for point-of-care diagnostics.Biosensors & bioelectronics · 2020Review
- The vision of point-of-care PCR tests for the COVID-19 pandemic and beyond.Trends in analytical chemistry : TRAC · 2020Review
- Article
- A strategy for preparing non-fluorescent graphene oxide quantum dots as fluorescence quenchers in quantitative real-time PCR.RSC advances · 2020Article
- Physical and chemical template-blocking strategies in the exponential amplification reaction of circulating microRNAs.Analytical and bioanalytical chemistry · 2020Article
- CRISPR-Cas12a based internal negative control for nonspecific products of exponential rolling circle amplification.Nucleic acids research · 2020Article
- Isothermal digital detection of microRNAs using background-free molecular circuit.Science advances · 2020Article
- Symmetric exponential amplification reaction-based DNA nanomachine for the fluorescent detection of nucleic acids.RSC advances · 2019Article
- rEXPAR: An Isothermal Amplification Scheme That Is Robust to Autocatalytic Parasites.Biochemistry · 2019Article
Corrections and comments
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Authors and funding
8 authors.
Funding
Abstract
Rapid isothermal nucleic acid amplification technologies can enable diagnosis of human pathogens and genetic variations in a simple, inexpensive, user-friendly format. The isothermal exponential amplification reaction (EXPAR) efficiently amplifies short oligonucleotides called triggers in less than 10 min by means of thermostable polymerase and nicking endonuclease activities. We recently demonstrated that this reaction can be coupled with upstream generation of trigger oligonucleotides from a genomic target sequence, and with downstream visual detection using DNA-functionalized gold nanospheres. The utility of EXPAR in clinical diagnostics is, however, limited by a nonspecific background amplification phenomenon, which is further investigated in this report. We found that nonspecific background amplification includes an early phase and a late phase. Observations related to late phase background amplification are in general agreement with literature reports of ab initio DNA synthesis. Early phase background amplification, which limits the sensitivity of EXPAR, differs however from previous reports of nonspecific DNA synthesis. It is observable in the presence of single-stranded oligonucleotides following the EXPAR template design rules and generates the trigger sequence expected for the EXPAR template present in the reaction. It appears to require interaction between the DNA polymerase and the single-stranded EXPAR template. Early phase background amplification can be suppressed or eliminated by physically separating the template and polymerase until the final reaction temperature has been reached, thereby enabling detection of attomolar starting trigger concentrations.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.