| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Molecular Diagnostics and Genetics |
1 Department of Microbiology, The University of Hong Kong, Hong Kong SAR.
2 Department of Microbiology, Queen Mary Hospital, Hong Kong SAR.
aAddress correspondence to this author at: Department of Microbiology, University of Hong Kong, Queen Mary Hospital, Pokfulam, Hong Kong SAR. Fax 852-28551241; e-mail llmpoon{at}hkucc.hku.hk.
Background: A novel coronavirus (CoV) was recently identified as the agent for severe acute respiratory syndrome (SARS). We compared the abilities of conventional and real-time reverse transcription-PCR (RT-PCR) assays to detect SARS CoV in clinical specimens.
Methods: RNA samples isolated from nasopharyngeal aspirate (NPA; n = 170) and stool (n = 44) were reverse-transcribed and tested by our in-house conventional RT-PCR assay. We selected 98 NPA and 37 stool samples collected at different times after the onset of disease and tested them in a real-time quantitative RT-PCR specific for the open reading frame (ORF) 1b region of SARS CoV. Detection rates for the conventional and real-time quantitative RT-PCR assays were compared. To investigate the nature of viral RNA molecules in these clinical samples, we determined copy numbers of ORF 1b and nucleocapsid (N) gene sequences of SARS CoV.
Results: The quantitative real-time RT-PCR assay was more sensitive than the conventional RT-PCR assay for detecting SARS CoV in samples collected early in the course of the disease. Real-time assays targeted at the ORF 1b region and the N gene revealed that copy numbers of ORF 1b and N gene sequences in clinical samples were similar.
Conclusions: NPA and stool samples can be used for early diagnosis of SARS. The real-time quantitative RT-PCR assay for SARS CoV is potentially useful for early detection of SARS CoV. Our results suggest that genomic RNA is the predominant viral RNA species in clinical samples.
The following articles in journals at HighWire Press have cited this article:
|
|
 |
|
  J. S. Sparks, E. F. Donaldson, X. Lu, R. S. Baric, and M. R. Denison A Novel Mutation in Murine Hepatitis Virus nsp5, the Viral 3C-Like Proteinase, Causes Temperature-Sensitive Defects in Viral Growth and Protein Processing J. Virol., June 15, 2008; 82(12): 5999 - 6008. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Fujimoto, K.-H. Chan, K. Takeda, K.-F. Lo, R. H. K. Leung, and T. Okamoto Sensitive and Specific Enzyme-Linked Immunosorbent Assay Using Chemiluminescence for Detection of Severe Acute Respiratory Syndrome Viral Infection J. Clin. Microbiol., January 1, 2008; 46(1): 302 - 310. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. H. Chan, K. Sonnenberg, M. Niedrig, S. Y. Lam, C. M. Pang, K. M. Chan, S. K. Ma, W. H. Seto, and J. S. M. Peiris Use of Antibody Avidity Assays for Diagnosis of Severe Acute Respiratory Syndrome Coronavirus Infection Clin. Vaccine Immunol., November 1, 2007; 14(11): 1433 - 1436. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. C. C. Cheng, S. K. P. Lau, P. C. Y. Woo, and K. Y. Yuen Severe Acute Respiratory Syndrome Coronavirus as an Agent of Emerging and Reemerging Infection Clin. Microbiol. Rev., October 1, 2007; 20(4): 660 - 694. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y.-P. Shih, C.-Y. Chen, S.-J. Liu, K.-H. Chen, Y.-M. Lee, Y.-C. Chao, and Y.-M. A. Chen Identifying Epitopes Responsible for Neutralizing Antibody and DC-SIGN Binding on the Spike Glycoprotein of the Severe Acute Respiratory Syndrome Coronavirus. J. Virol., November 1, 2006; 80(21): 10315 - 10324. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Louie, A. E. Simor, S. Chong, K. Luinstra, A. Petrich, J. Mahony, M. Smieja, G. Johnson, F. Gharabaghi, R. Tellier, et al. Detection of Severe Acute Respiratory Syndrome Coronavirus in Stool Specimens by Commercially Available Real-Time Reverse Transcriptase PCR Assays J. Clin. Microbiol., November 1, 2006; 44(11): 4193 - 4196. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Kuypers, N. Wright, J. Ferrenberg, M.-L. Huang, A. Cent, L. Corey, and R. Morrow Comparison of real-time PCR assays with fluorescent-antibody assays for diagnosis of respiratory virus infections in children. J. Clin. Microbiol., July 1, 2006; 44(7): 2382 - 2388. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. J. Espy, J. R. Uhl, L. M. Sloan, S. P. Buckwalter, M. F. Jones, E. A. Vetter, J. D. C. Yao, N. L. Wengenack, J. E. Rosenblatt, F. R. Cockerill III, et al. Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing Clin. Microbiol. Rev., January 1, 2006; 19(1): 165 - 256. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. P. Yip, S. S. T. To, P. H.M. Leung, T. S. Cheung, P. K.C. Cheng, and W. W.L. Lim Use of Dual TaqMan Probes to Increase the Sensitivity of 1-Step Quantitative Reverse Transcription-PCR: Application to the Detection of SARS Coronavirus Clin. Chem., October 1, 2005; 51(10): 1885 - 1888. [Full Text] [PDF]
|
|
|
|
 |
|
 P. L. Ho, P. H. Chau, P. S. F. Yip, G. C. Ooi, P. L. Khong, J. C. Ho, P. C. Wong, C. Ko, C. Yan, and K. W. Tsang A prediction rule for clinical diagnosis of severe acute respiratory syndrome Eur. Respir. J., September 1, 2005; 26(3): 474 - 479. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. L. M. Poon, B. W. Y. Wong, K. H. Chan, S. S. F. Ng, K. Y. Yuen, Y. Guan, and J. S. M. Peiris Evaluation of Real-Time Reverse Transcriptase PCR and Real-Time Loop-Mediated Amplification Assays for Severe Acute Respiratory Syndrome Coronavirus Detection J. Clin. Microbiol., July 1, 2005; 43(7): 3457 - 3459. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Y. Cheung, L. L. M. Poon, I. H. Y. Ng, W. Luk, S.-F. Sia, M. H. S. Wu, K.-H. Chan, K.-Y. Yuen, S. Gordon, Y. Guan, et al. Cytokine Responses in Severe Acute Respiratory Syndrome Coronavirus-Infected Macrophages In Vitro: Possible Relevance to Pathogenesis J. Virol., June 15, 2005; 79(12): 7819 - 7826. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. Hu, B. Bai, Z. Hu, Z. Chen, X. An, L. Tang, J. Yang, H. Wang, and H. Wang 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., May 1, 2005; 43(5): 2041 - 2046. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Di, W. Hao, Y. Gao, M. Wang, Y.-d. Wang, L.-w. Qiu, K. Wen, D.-h. Zhou, X.-w. Wu, E.-j. Lu, et al. Monoclonal Antibody-Based Antigen Capture Enzyme-Linked Immunosorbent Assay Reveals High Sensitivity of the Nucleocapsid Protein in Acute-Phase Sera of Severe Acute Respiratory Syndrome Patients Clin. Vaccine Immunol., January 1, 2005; 12(1): 135 - 140. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K.W. Tsang, G.C. Ooi, and P.L. Ho Diagnosis and pharmacotherapy of severe acute respiratory syndrome: what have we learnt? Eur. Respir. J., December 1, 2004; 24(6): 1025 - 1032. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. L.M. Poon, C. S.W. Leung, M. Tashiro, K. H. Chan, B. W.Y. Wong, K. Y. Yuen, Y. Guan, and J. S.M. Peiris Rapid Detection of the Severe Acute Respiratory Syndrome (SARS) Coronavirus by a Loop-Mediated Isothermal Amplification Assay Clin. Chem., June 1, 2004; 50(6): 1050 - 1052. [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
