Quantitation of severe acute respiratory syndrome coronavirus genome by real-time polymerase chain reaction assay using minor groove binder DNA probe technology

Hsi Hsun Lin, Shiow J. Wang, Yung Ching Liu, S. S J Lee, Chun K. Hwang, Yao Shen Chen, Shue R. Wann, Yi L. Shih

研究成果: 雜誌貢獻文章同行評審

5 引文 斯高帕斯(Scopus)

摘要

The ability to rapidly recognize severe acute respiratory syndrome coronavirus (SARS-CoV) as a cause of infections is critical to quickly limiting further spread of the disease. A rapid, sensitive, and specific laboratory diagnostic test is needed to confirm outbreaks of SARS-CoV infection and distinguish it from other diseases that can cause similar clinical symptoms. An improved TaqMan technology using minor groove binder (MGB) probes was used to detect and quantify SARS-CoV in suspected patients. SARS-CoV primers and probe were designed based on the open reading frame 1b sequence, which encodes coronavirus replicase protein. A linear standard curve with R2 > 0.99 was obtained, and the threshold sensitivity was 10 genome equivalents per reaction. Interassay coefficients of variation were 1.73 to 2.72%, indicating good reproducibility of this method. A total of 228 specimens from 151 suspected patients were quantified by this method, 13 specimens from 6 patients were positive with viral load range from 362 to 36,240,000 genome equivalents/mL. In conclusion, the high sensitivity and reproducibility of the real-time polymerase chain reaction SARS-CoV RNA quantitation using MGB probe allowed the screening of large numbers of clinical samples.

原文英語
頁(從 - 到)258-265
頁數8
期刊Journal of Microbiology, Immunology and Infection
37
發行號5
出版狀態已發佈 - 十月 2004
對外發佈

ASJC Scopus subject areas

  • 免疫學與微生物學 (全部)
  • 免疫學和過敏
  • 微生物學(醫學)

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