The DOI matches its Crossref record; 26 of the 26 references it cites were found in a registry or an index.
Real-Time Reverse Transcription Loop-Mediated Isothermal Amplification for Rapid Detection of West Nile Virus
Open accessAbstract
A one-step, single tube, real-time accelerated reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detecting the envelope gene of West Nile (WN) virus. The RT-LAMP assay is a novel method of gene amplification that amplifies nucleic acid with high specificity, efficiency, and rapidity under isothermal conditions with a set of six specially designed primers that recognize eight distinct sequences of the target. The whole procedure is very simple and rapid, and amplification can be obtained in less than 1 h by incubating all of the reagents in a single tube with reverse transcriptase and Bst DNA polymerase at 63°C. Detection of gene amplification could be accomplished by agarose gel electrophoresis, as well as by real-time monitoring in an inexpensive turbidimeter. When the sensitivity of the RT-LAMP assay was compared to that of conventional RT-PCR, it was found that the RT-LAMP assay demonstrated 10-fold higher sensitivity compared to RT-PCR, with a detection limit of 0.1 PFU of virus. By using real-time monitoring, 10 4 PFU of virus could be detected in as little as 17 min. The specificity of the RT-LAMP assay was validated by the absence of any cross-reaction with other, closely related, members of the Flavivirus group, followed by restriction digestion and nucleotide sequencing of the amplified product. These results indicate that the RT-LAMP assay is extremely rapid, cost-effective, highly sensitive, and specific and has potential usefulness for rapid, comprehensive WN virus surveillance along with virus isolation and/or serology.
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Cites
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Cited by
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Current and Developing Technologies for Monitoring Agents of Bioterrorism and Biowarfare
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Augustine, R., Hasan, A., Das, S., Ahmed, R., Mori, Y., Notomi, T., Kevadiya, B., & Thakor, A. (2020) · Biology · doi:10.3390/biology9080182 · Cited by 253
Real-Time Fluorescence Loop Mediated Isothermal Amplification for the Diagnosis of Malaria
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Self-citation
Metadata
- Authors
- Parida, M., Posadas, G., Inoue, S., Hasebe, F., & Morita, K.
- Venue
- Journal of Clinical Microbiology
- Publisher
- American Society for Microbiology
- Published
- 2004-01
- Volume
- 42
- Issue
- 1
- Pages
- 257-263
- ISSN
- 0095-1137 (ISSN-L)
- 1098-660X
- OpenAlex
- W2149136689
- Cited by
- 438
- References
- 26
- FWCI
- 14.64
- Type
- journal article
- Language
- en
- Open access
- green
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- Parida, M., Posadas, G., Inoue, S., Hasebe, F., & Morita, K. (2004). Real-Time Reverse Transcription Loop-Mediated Isothermal Amplification for Rapid Detection of West Nile Virus. Journal of Clinical Microbiology. https://doi.org/10.1128/jcm.42.1.257-263.2004
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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