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Helicase‐dependent isothermal DNA amplification
Open accessAbstract
Polymerase chain reaction is the most widely used method for in vitro DNA amplification. However, it requires thermocycling to separate two DNA strands. In vivo , DNA is replicated by DNA polymerases with various accessory proteins, including a DNA helicase that acts to separate duplex DNA. We have devised a new in vitro isothermal DNA amplification method by mimicking this in vivo mechanism. Helicase‐dependent amplification (HDA) utilizes a DNA helicase to generate single‐stranded templates for primer hybridization and subsequent primer extension by a DNA polymerase. HDA does not require thermocycling. In addition, it offers several advantages over other isothermal DNA amplification methods by having a simple reaction scheme and being a true isothermal reaction that can be performed at one temperature for the entire process. These properties offer a great potential for the development of simple portable DNA diagnostic devices to be used in the field and at the point‐of‐care.
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Citations
Cites
Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA Polymerase
Saiki, R. K., Gelfand, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. T., Mullis, K. B., & Erlich, H. A. (1988) · Science · doi:10.1126/science.2448875 · Cited by 17,719
Cited by
Kaminski, M. M., Abudayyeh, O. O., Gootenberg, J. S., Zhang, F., & Collins, J. J. (2021) · Nature Biomedical Engineering · doi:10.1038/s41551-021-00760-7 · Cited by 1,259
Quan, P.-L., Sauzade, M., & Brouzes, E. (2018) · Sensors · doi:10.3390/s18041271 · Cited by 664
Methods in virus diagnostics: From ELISA to next generation sequencing
Boonham, N., Kreuze, J., Winter, S., van der Vlugt, R., Bergervoet, J., Tomlinson, J., & Mumford, R. (2014) · Virus Research · doi:10.1016/j.virusres.2013.12.007 · Cited by 448
Shen, F., Du, W., Kreutz, J. E., Fok, A., & Ismagilov, R. F. (2010) · Lab on a Chip · doi:10.1039/c004521g · Cited by 295
Shen, F., Davydova, E. K., Du, W., Kreutz, J. E., Piepenburg, O., & Ismagilov, R. F. (2011) · Analytical Chemistry · doi:10.1021/ac200247e · Cited by 240
Metadata
- Authors
- Vincent, M., Xu, Y., & Kong, H.
- Venue
- EMBO reports
- Published
- 2004-08
- Volume
- 5
- Issue
- 8
- Pages
- 795-800
- ISSN
- 1469-221X (ISSN-L)
- 1469-3178
- OpenAlex
- W2117240806
- Cited by
- 877
- References
- 1
- FWCI
- 5.05
- Type
- journal article
- Language
- en
- Open access
- bronze
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
