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Journal of Clinical Microbiology, October 2003, p. 4521-4524, Vol. 41, No. 10
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.10.4521-4524.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Evaluation of Reverse Transcription-PCR Assays for Rapid Diagnosis of Severe Acute Respiratory Syndrome Associated with a Novel Coronavirus

W. C. Yam, K. H. Chan, L. L. M. Poon, Y. Guan, K. Y. Yuen, W. H. Seto, and J. S. M. Peiris*

Department of Microbiology, Queen Mary Hospital, The University of Hong Kong, Hong Kong, People's Republic of China

Received 15 May 2003/ Returned for modification 17 June 2003/ Accepted 21 July 2003

The reverse transcription (RT)-PCR protocols of two World Health Organization (WHO) severe acute respiratory syndrome (SARS) network laboratories (WHO SARS network laboratories at The University of Hong Kong [WHO-HKU] and at the Bernhard-Nocht Institute in Hamburg, Germany [WHO-Hamburg]) were evaluated for rapid diagnosis of a novel coronavirus (CoV) associated with SARS in Hong Kong. A total of 303 clinical specimens were collected from 163 patients suspected to have SARS. The end point of both WHO-HKU and WHO-Hamburg RT-PCR assays was determined to be 0.1 50% tissue culture infective dose. Using seroconversion to CoV as the "gold standard" for SARS CoV diagnosis, WHO-HKU and WHO-Hamburg RT-PCR assays exhibited diagnostic sensitivities of 61 and 68% (nasopharyngeal aspirate specimens), 65 and 72% (throat swab specimens), 50 and 54% (urine specimens), and 58 and 63% (stool specimens), respectively, with an overall specificity of 100%. For patients confirmed to have SARS CoV and from whom two or more respiratory specimens were collected, testing the second specimen increased the sensitivity from 64 and 71% to 75 and 79% for the WHO-HKU and WHO-Hamburg RT-PCR assays, respectively. Testing more than one respiratory specimen will maximize the sensitivity of PCR assays for SARS CoV.

* Corresponding author. Mailing address: Department of Microbiology, The University of Hong Kong, University Pathology Building, Queen Mary Hospital Compound, Pokfulam, Hong Kong SAR, People's Republic of China. Phone: 852-28554888. Fax: 852-28551241. E-mail: malik{at}hku.hk.

Journal of Clinical Microbiology, October 2003, p. 4521-4524, Vol. 41, No. 10
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.10.4521-4524.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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