title: Development of loop-mediated isothermal amplification assay for detection of human coronavirus-NL63.
abstract: 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.
authors.affiliation: Microbiology Department, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Krakow, Poland. k.a.pyrc@uj.edu.pl
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Pyrc
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Milewska
authors.forename: Jan
authors.initials: J
authors.lastname: Potempa
mesh_terms: D018352:Coronavirus Infections / Q000821:virology; D058957:Coronavirus NL63, Human / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*