Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 175, Issue 1, Pages 133-136Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2011.04.024
Keywords
Coronavirus; HCoV-NL63; LAMP; Loop-mediated isothermal amplification
Funding
- Ministry of Scientific Research, Poland [0095/B/P01/2009/37]
- Ministry of Science and Higher Education, Poland [IP 2010 033870]
- Jagiellonian University [DS/9/WBBiB]
- Department of Scientific Research, Polish Ministry of Science and Education [1642/B/P01/2008/35]
- Foundation for Polish Science [DPS/424-329/10]
- European Union [POIG.02.01.00-12-064/08]
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Human coronavirus NL63 was identified in 2004 in the Netherlands. Due to the high prevalence and world-wide distribution of this pathogen, it is essential to develop a sensitive and specific detection assay suitable for use in a routine diagnostic laboratory. Techniques based on PCR or real-time PCR are laborious and expensive. Detailed analysis of the HCoV-NL63 genome permitted the identification of a conserved nucleic acid sequential motif, which was sufficient for the design of a loop-mediated isothermal amplification (LAMP) assay. Evaluation of the method showed that the test is specific to HCoV-NL63 and that it does not cross-react with other respiratory viruses. The detection limit was found to be 1 copy of RNA template per reaction in cell culture supernatants and clinical specimens. (C) 2011 Elsevier B.V. All rights reserved.
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