Archives of Virology

, Volume 152, Issue 1, pp 41–58

SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses

  • S. Escutenaire
  • N. Mohamed
  • M. Isaksson
  • P. Thorén
  • B. Klingeborn
  • S. Belák
  • M. Berg
  • J. Blomberg
Article
  • 560 Downloads

Summary.

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.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • S. Escutenaire
    • 1
  • N. Mohamed
    • 2
  • M. Isaksson
    • 1
  • P. Thorén
    • 1
  • B. Klingeborn
    • 1
  • S. Belák
    • 1
    • 3
  • M. Berg
    • 3
  • J. Blomberg
    • 2
  1. 1.Department of VirologyNational Veterinary InstituteUppsalaSweden
  2. 2.Department of Medical Sciences, Section of Clinical Virology, Academic HospitalUppsala UniversityUppsalaSweden
  3. 3.Department of Biomedical Sciences and Veterinary Public Health, Section of Parasitology and VirologySwedish University of Agricultural SciencesUppsalaSweden

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