Virologica Sinica

, Volume 24, Issue 3, pp 187–193

Evaluation of sensitivities and specificities of SARS-CoV detection by real-time quantitative reverse transcription-PCR assays

  • Li-li Xu
  • Zhi-hong Hu
  • Hua-lin Wang
  • Xiao Han
  • Fei Deng
Article

DOI: 10.1007/s12250-009-3021-8

Cite this article as:
Xu, L., Hu, Z., Wang, H. et al. Virol. Sin. (2009) 24: 187. doi:10.1007/s12250-009-3021-8
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Abstract

The etiological agent of severe acute respiratory syndrome (SARS) was identified as a new coronavirus, termed SARS-CoV. Establishment of an efficient and sensitive diagnostic system of SARS-CoV genetic materials is crucial for SARS control. In this study, we quantified SARS-CoV mRNAs in both infected cell culture lysate and in supernatant by using Real-time quantitative revere transcription-PCR based on EvaGreen™ dye and Taqman-MGB probes. For extensive evaluation of sensitivities and specificities, 13 pairs of primers and 4 probes were designed based on different genes of SARS-CoV. Glyceraldehydes-3-phosphate dehydrogenase (GAPDH) was selected as the internal control gene. Results showed that S-gene-specific PCR was the most sensitive for detection, but because of its sequence variability in the different viral strains, primers and a probe based on the N gene were suitable substitutions. Meanwhile, we found the mRNA concentrations in cell culture lysates were much higher than in cell supernatant and facilited more sensitive detection of the SARS-CoV.

Key words

SARS-CoV Sensitivities Specificities Evaluation 

CLC number

R373 

Copyright information

© Wuhan Institute of Virology, CAS and Springer-Verlag GmbH 2009

Authors and Affiliations

  • Li-li Xu
    • 1
  • Zhi-hong Hu
    • 1
  • Hua-lin Wang
    • 1
  • Xiao Han
    • 1
  • Fei Deng
    • 1
  1. 1.State Key Laboratory of Virology, Wuhan Institute of VirologyChinese Academy of SciencesWuhanChina