This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Bressler, A. M. Articles by Nolte, F. S. Search for Related Content PubMed PubMed Citation Articles by Bressler, A. M. Articles by Nolte, F. S.

Preclinical Evaluation of Two Real-Time, Reverse Transcription-PCR Assays for Detection of the Severe Acute Respiratory Syndrome Coronavirus

Departments of Medicine (Infectious Diseases) and Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia

Received 23 September 2003/ Returned for modification 27 October 2003/ Accepted 14 November 2003

We verified the analytical performance characteristics of a previously described real-time reverse transcription-PCR (RT-PCR) assay targeting the open reading frame (ORF) 1b region of the severe acute respiratory syndrome coronavirus (SARS-CoV) with RNA transcripts. We then compared it to a novel nucleocapsid gene real-time RT-PCR assay with genomic RNA. The assays differed only in the primer and probe sequences and final concentrations. A commercially available armored RNA (Ambion, Austin, Tex.) was evaluated as positive control for the ORF 1b assay. The analytical sensitivity, reproducibility, amplification efficiency, and dynamic range of the assays were similar. Both were specific for SARS-CoV as determined by testing against human CoV 229E and OC43, specimens from patients without SARS, and by BLAST searches of GenBank for primer and probe sequence homology. The armored RNA was found to be a suitable positive control for the ORF 1b assay that could be reliably recovered and amplified from a variety of clinical specimens.

This article has been cited by other articles:

• Wei, Y., Yang, C., Wei, B., Huang, J., Wang, L., Meng, S., Zhang, R., Li, J. (2008). RNase-Resistant Virus-Like Particles Containing Long Chimeric RNA Sequences Produced by Two-Plasmid Coexpression System. J. Clin. Microbiol. 46: 1734-1740 [Abstract] [Full Text]  
• Yu, X.-F., Pan, J.-C., Ye, R., Xiang, H.-Q., Kou, Y., Huang, Z.-C. (2008). Preparation of Armored RNA as a Control for Multiplex Real-Time Reverse Transcription-PCR Detection of Influenza Virus and Severe Acute Respiratory Syndrome Coronavirus. J. Clin. Microbiol. 46: 837-841 [Abstract] [Full Text]  
• Cheng, Y., Niu, J., Zhang, Y., Huang, J., Li, Q. (2006). Preparation of His-Tagged Armored RNA Phage Particles as a Control for Real-Time Reverse Transcription-PCR Detection of Severe Acute Respiratory Syndrome Coronavirus.. J. Clin. Microbiol. 44: 3557-3561 [Abstract] [Full Text]  
• Zhu, H., Hu, S., Jona, G., Zhu, X., Kreiswirth, N., Willey, B. M., Mazzulli, T., Liu, G., Song, Q., Chen, P., Cameron, M., Tyler, A., Wang, J., Wen, J., Chen, W., Compton, S., Snyder, M. (2006). Severe acute respiratory syndrome diagnostics using a coronavirus protein microarray.. Proc. Natl. Acad. Sci. USA 103: 4011-4016 [Abstract] [Full Text]  
• Hietala, S. K., Crossley, B. M. (2006). Armored RNA as Virus Surrogate in a Real-Time Reverse Transcriptase PCR Assay Proficiency Panel. J. Clin. Microbiol. 44: 67-70 [Abstract] [Full Text]  
• Espy, M. J., Uhl, J. R., Sloan, L. M., Buckwalter, S. P., Jones, M. F., Vetter, E. A., Yao, J. D. C., Wengenack, N. L., Rosenblatt, J. E., Cockerill, F. R. III, Smith, T. F. (2006). Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing. Clin. Microbiol. Rev. 19: 165-256 [Abstract] [Full Text]  
• Mahony, J. B., Richardson, S. (2005). Molecular Diagnosis of Severe Acute Respiratory Syndrome: The State of the Art. J. Mol. Diagn. 7: 551-559 [Abstract] [Full Text]  
• Hu, W., Bai, B., Hu, Z., Chen, Z., An, X., Tang, L., Yang, J., Wang, H., Wang, H. (2005). Development and Evaluation of a Multitarget Real-Time Taqman Reverse Transcription-PCR Assay for Detection of the Severe Acute Respiratory Syndrome-Associated Coronavirus and Surveillance for an Apparently Related Coronavirus Found in Masked Palm Civets. J. Clin. Microbiol. 43: 2041-2046 [Abstract] [Full Text]  
• Liu, I-J., Chen, P.-J., Yeh, S.-H., Chiang, Y.-P., Huang, L.-M., Chang, M.-F., Chen, S.-Y., Yang, P.-C., Chang, S.-C., Wang, W.-K., the SARS Research Group of the National Taiwan Uni, (2005). Immunofluorescence Assay for Detection of the Nucleocapsid Antigen of the Severe Acute Respiratory Syndrome (SARS)-Associated Coronavirus in Cells Derived from Throat Wash Samples of Patients with SARS. J. Clin. Microbiol. 43: 2444-2448 [Abstract] [Full Text]  
• WalkerPeach, C. R., Pasloske, B. L. (2004). DNA Bacteriophage as Controls for Clinical Viral Testing. Clin. Chem. 50: 1970-1971 [Full Text]