Development of loop-mediated isothermal amplification assay for detection of human coronavirus-NL63

J Virol Methods. 2011 Jul;175(1):133-6. doi: 10.1016/j.jviromet.2011.04.024. Epub 2011 Apr 27.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Coronavirus Infections / virology
  • Coronavirus NL63, Human / genetics*
  • Coronavirus NL63, Human / isolation & purification
  • Humans
  • Nucleic Acid Amplification Techniques / methods*