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.
【저자키워드】 coronavirus, Loop-mediated isothermal amplification, LAMP, HCoV-NL63, 【초록키워드】 coronavirus, Human, Genome, diagnostic, Laboratory, RNA, Prevalence, nucleic acid, PCR, pathogen, Real-time PCR, isothermal amplification, NL63, distribution, development, specimens, HCoV-NL63, Detection limit, Analysis, Other respiratory viruses, Netherlands, technique, motif, cell culture supernatant, found, develop, conserved, cross-react,