Comprehensive detection and identification of human coronaviruses, including the SARS-associated coronavirus, with a single RT-PCR assay

D Adachi; G Johnson; R Draker; M Ayers; T Mazzulli; P J Talbot; R Tellier

doi:10.1016/j.jviromet.2004.07.008

Comprehensive detection and identification of human coronaviruses, including the SARS-associated coronavirus, with a single RT-PCR assay

J Virol Methods. 2004 Dec 1;122(1):29-36. doi: 10.1016/j.jviromet.2004.07.008.

Authors

D Adachi¹, G Johnson, R Draker, M Ayers, T Mazzulli, P J Talbot, R Tellier

Affiliation

¹ Division of Microbiology, The Hospital for Sick Children, 555 University Avenue, Toronto, Ont., Canada M5G 1X8.

Abstract

The SARS-associated human coronavirus (SARS-HCoV) is a newly described, emerging virus conclusively established as the etiologic agent of the severe acute respiratory syndrome (SARS). This study presents a single-tube RT-PCR assay that can detect with high analytical sensitivity the SARS-HCoV, as well as several other coronaviruses including other known human respiratory coronaviruses (HCoV-OC43 and HCoV-229E). Species identification is provided by sequencing the amplicon, although a rapid screening test by restriction enzyme analysis has proved to be very useful for the analysis of samples obtained during the SARS outbreak in Toronto, Canada.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Base Sequence
Coronavirus / classification*
Coronavirus / genetics
Coronavirus / isolation & purification*
Coronavirus 229E, Human / classification
Coronavirus 229E, Human / genetics
Coronavirus 229E, Human / isolation & purification
Coronavirus Infections / diagnosis*
Coronavirus Infections / virology
Coronavirus OC43, Human / classification
Coronavirus OC43, Human / genetics
Coronavirus OC43, Human / isolation & purification
DNA, Complementary
DNA, Viral / analysis
DNA, Viral / isolation & purification
Deoxyribonucleases, Type II Site-Specific / metabolism
Humans
Molecular Sequence Data
Phylogeny
Reverse Transcriptase Polymerase Chain Reaction / methods*
Sensitivity and Specificity
Sequence Alignment
Sequence Analysis, DNA
Severe Acute Respiratory Syndrome / diagnosis*
Severe Acute Respiratory Syndrome / virology
Severe acute respiratory syndrome-related coronavirus / classification
Severe acute respiratory syndrome-related coronavirus / genetics
Severe acute respiratory syndrome-related coronavirus / isolation & purification*

Substances

DNA, Complementary
DNA, Viral
endodeoxyribonuclease AluI
Deoxyribonucleases, Type II Site-Specific