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Journal of Clinical Microbiology, May 2005, p. 2339-2344, Vol. 43, No. 5
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.5.2339-2344.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Rapid and Sensitive Detection of Severe Acute Respiratory Syndrome Coronavirus by Rolling Circle Amplification

Bin Wang,1* Simon J. Potter,1 Yiguang Lin,2 Anthony L. Cunningham,1 Dominic E. Dwyer,1,3 Yuelong Su,4 Xuejun Ma,4 Yunde Hou,4 and Nitin K. Saksena1

Centre for Virus Research, Westmead Millennium Institute, The University of Sydney,1 Department of Health Sciences, University of Technology,2 CIDMLS, ICPMR, Westmead Hospital, Sydney, New South Wales, Australia,3 Beijing JDK Bio-technology Institute, National Institute of Virology, Chinese Academy of Preventive Medicine, Beijing, China4

Received 28 October 2004/ Returned for modification 17 November 2004/ Accepted 5 January 2005

The severe acute respiratory syndrome (SARS) epidemic of 2003 was responsible for 774 deaths and caused significant economic damage worldwide. Since July 2003, a number of SARS cases have occurred in China, raising the possibility of future epidemics. We describe here a rapid, sensitive, and highly efficient assay for the detection of SARS coronavirus (SARS-CoV) in cultured material and a small number (n = 7) of clinical samples. Using rolling circle amplification (RCA), we were able to achieve sensitive detection levels of SARS-CoV RNA in both solid and liquid phases. The main advantage of RCA is that it can be performed under isothermal conditions with minimal reagents and avoids the generation of false-positive results, a problem that is frequently encountered in PCR-based assays. Furthermore, the RCA technology provides a faster, more sensitive, and economical option to currently available PCR-based methods.

* Corresponding author. Mailing address: Retroviral Genetics Laboratory, Centre for Virus Research, Westmead Millennium Institute, The University of Sydney, Darcy Rd., Westmead, Sydney, NSW 2145, Australia. Phone: 61-2-98459107. Fax: 61-2-98459103. E-mail: bin_wang{at}wmi.usyd.edu.au.

Journal of Clinical Microbiology, May 2005, p. 2339-2344, Vol. 43, No. 5
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.5.2339-2344.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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