Coupling multiplex RT-PCR to a gene chip assay for sensitive and semiquantitative detection of severe acute respiratory syndrome-coronavirus

Lab Invest. 2004 Sep;84(9):1085-91. doi: 10.1038/labinvest.3700136.

Abstract

An early and accurate diagnostic assay for severe acute respiratory syndrome (SARS) is crucial for infection control. However, most of the diagnostic methods available today, such as real-time reverse transcriptase-polymerase chain reaction (RT-PCR), require a second detection method for confirmation because they detect a single sequence region of the SARS-coronavirus (SARS-CoV). For sensitive and accurate early diagnosis, we report a novel assay system combining multiplex RT-PCR and a diagnostic gene chip to detect multiple virus-specific genomic sequences of SARS-CoV. With 53 clinical specimens, we successfully demonstrate that this technique offers not only a high-accuracy diagnosis for early SARS infection but also a semiquantitative assay.

MeSH terms

  • DNA Primers / chemistry
  • Humans
  • Oligonucleotide Array Sequence Analysis / methods*
  • RNA, Complementary / chemistry
  • RNA, Viral / analysis*
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction*
  • 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 Primers
  • RNA, Complementary
  • RNA, Viral