SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses
2007 (English)In: Archives of Virology, ISSN 0304-8608, E-ISSN 1432-8798, Vol. 152, no 1, 41-58 p.Article in journal (Refereed) Published
Coronaviruses are etiologic agents of respiratory and enteric diseases in humans and in animals. In this study, a one-step real-time reverse transcription-polymerase chain reaction (RT-PCR) assay based on SYBR Green chemistry and degenerate primers was developed for the generic detection of coronaviruses. The primers, designed in the open reading frame 1b, enabled the detection of 32 animal coronaviruses including strains of canine coronavirus, feline coronavirus, transmissible gastroenteritis virus (TGEV), bovine coronavirus (BCoV), murine hepatitis virus (MHV) and infectious bronchitis virus (IBV). A specific amplification was also observed with the human coronaviruses (HCoV) HCoV-NL63, HCoV-OC43, HCoV-229E and severe acute respiratory syndrome coronavirus (SARS-CoV). The real-time RT-PCR detected down to 10 cRNA copies from TGEV, BCoV, SARS-CoV and IBV. In addition, the assay exhibited a high sensitivity and specificity on clinical samples from different animal species. The developed assay represents a potential tool for laboratory diagnostics and for detecting still uncharacterized coronaviruses.
Place, publisher, year, edition, pages
2007. Vol. 152, no 1, 41-58 p.
Medical and Health Sciences
IdentifiersURN: urn:nbn:se:uu:diva-11882DOI: 10.1007/s00705-006-0840-xISI: 000242998400004PubMedID: 16941059OAI: oai:DiVA.org:uu-11882DiVA: diva2:39651