The design and application of DNA chips for early detection of SARS-CoV from clinical samples

J Clin Virol. 2005 Jun;33(2):123-31. doi: 10.1016/j.jcv.2004.10.019. Epub 2005 Jan 19.

Abstract

Background: SARS coronavirus has been identified as the cause of severe acute respiratory syndrome (SARS). Few tests allow confirmation or exclusion of SARS within the first few days of infection. A gene chip is a useful tool for the study of microbial infections mainly for its capability of performing multi-target analysis in a single test.

Objectives: Investigate the possibility of early detection of SARS virus from clinical samples using the gene chip-based method.

Study design: We purified RNA from SARS-CoV obtained from routinely collected peripheral blood and sputum samples of 34 patients who had been identified as probable SARS patients by following the interim U.S. case definition. Four segments of the SARS-CoV were amplified using reverse transcription-nested PCR and the products examined using the 70-mer gene chips for SARS-CoV detection.

Results: A blind-test of both peripheral blood and sputum specimens lead to the positive detection of SARS-CoV in 31 out of 34 patients. SARS-CoV was not found in peripheral blood or sputum specimens from three patients. Two of the 34 patients were only 3 days post-onset of symptoms and were subsequently confirmed to be SARS positive. Our results indicate that the gene chip-based molecular test is specific for SARS-CoV and allows early detection of patients with SARS with detection rate about 8% higher than the single PCR test when the sputum sample is available.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood / virology
  • China
  • Early Diagnosis
  • Humans
  • Molecular Diagnostic Techniques*
  • Oligonucleotide Array Sequence Analysis*
  • Polymerase Chain Reaction
  • RNA, Viral / analysis
  • RNA, Viral / isolation & purification
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sensitivity and Specificity
  • Severe Acute Respiratory Syndrome / diagnosis*
  • Severe Acute Respiratory Syndrome / genetics
  • Severe acute respiratory syndrome-related coronavirus / genetics*
  • Severe acute respiratory syndrome-related coronavirus / isolation & purification*
  • Sputum / virology

Substances

  • RNA, Viral