A one step quantitative RT-PCR for detection of SARS coronavirus with an internal control for PCR inhibitors

J Clin Virol. 2004 Jul;30(3):214-7. doi: 10.1016/j.jcv.2003.12.007.

Abstract

In this study, we report a one step quantitative RT-PCR assay for severe acute respiratory syndrome (SARS) diagnosis. The overall detection rate for clinical samples collected from Days 1 to 9 of disease onset is 86.2% and the specificity of the assay is 100%. To detect false negative results due to PCR inhibitors or faulty RNA extraction, we have further modified this one step RT-PCR assay for simultaneous detection of severe acute respiratory syndrome (SARS) coronavirus (CoV) and 18S ribosomal RNA (rRNA) as an internal positive control.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • DNA, Ribosomal / analysis
  • Humans
  • Nasopharynx / virology
  • RNA, Ribosomal, 18S / genetics
  • RNA, Viral / analysis
  • RNA, Viral / isolation & purification
  • Reference Standards
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Severe Acute Respiratory Syndrome / diagnosis*
  • Severe Acute Respiratory Syndrome / virology
  • Severe acute respiratory syndrome-related coronavirus / genetics
  • Severe acute respiratory syndrome-related coronavirus / isolation & purification*

Substances

  • DNA, Ribosomal
  • RNA, Ribosomal, 18S
  • RNA, Viral