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

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)41-58
Number of pages18
JournalArchives of Virology
Volume152
Issue number1
DOIs
Publication statusPublished - 1 Jan 2007

Fingerprint

Coronavirus
Reverse Transcription
Polymerase Chain Reaction
Bovine Coronavirus
Transmissible gastroenteritis virus
Infectious bronchitis virus
Severe Acute Respiratory Syndrome
Human Coronavirus NL63
Human Coronavirus OC43
Canine Coronavirus
Human Coronavirus 229E
Feline Coronavirus
Murine hepatitis virus
Complementary RNA
Open Reading Frames
Sensitivity and Specificity

Cite this

Escutenaire, S. ; Mohamed, N. ; Isaksson, M. ; Thorén, P. ; Klingeborn, B. ; Belák, S. ; Berg, M. ; Blomberg, J. / SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses. In: Archives of Virology. 2007 ; Vol. 152, No. 1. pp. 41-58.
@article{f177c2b5130a472583e62524840a7581,
title = "SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses",
abstract = "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.",
author = "S. Escutenaire and N. Mohamed and M. Isaksson and P. Thor{\'e}n and B. Klingeborn and S. Bel{\'a}k and M. Berg and J. Blomberg",
year = "2007",
month = "1",
day = "1",
doi = "10.1007/s00705-006-0840-x",
language = "English",
volume = "152",
pages = "41--58",
journal = "Archives of Virology",
issn = "0304-8608",
publisher = "Springer-Verlag Wien",
number = "1",

}

Escutenaire, S, Mohamed, N, Isaksson, M, Thorén, P, Klingeborn, B, Belák, S, Berg, M & Blomberg, J 2007, 'SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses', Archives of Virology, vol. 152, no. 1, pp. 41-58. https://doi.org/10.1007/s00705-006-0840-x

SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses. / Escutenaire, S.; Mohamed, N.; Isaksson, M.; Thorén, P.; Klingeborn, B.; Belák, S.; Berg, M.; Blomberg, J.

In: Archives of Virology, Vol. 152, No. 1, 01.01.2007, p. 41-58.

Research output: Contribution to journalArticle

TY - JOUR

T1 - SYBR Green real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses

AU - Escutenaire, S.

AU - Mohamed, N.

AU - Isaksson, M.

AU - Thorén, P.

AU - Klingeborn, B.

AU - Belák, S.

AU - Berg, M.

AU - Blomberg, J.

PY - 2007/1/1

Y1 - 2007/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=33845785362&partnerID=8YFLogxK

U2 - 10.1007/s00705-006-0840-x

DO - 10.1007/s00705-006-0840-x

M3 - Article

C2 - 16941059

AN - SCOPUS:33845785362

VL - 152

SP - 41

EP - 58

JO - Archives of Virology

JF - Archives of Virology

SN - 0304-8608

IS - 1

ER -