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学科主题: 基础医学
题名:
A real-time PCR for SARS-coronavirus incorporating target gene pre-amplification
作者: Lau, LT; Fung, YWW; Wong, FPF; Lin, SSW; Wang, CR; Li, HL; Dillon, N; Collins, RA; Tam, JSL; Chan, PKS; Wang, CG; Yu, ACH
关键词: TaqMan real-time fluorescent polymerase chain reaction ; PCR ; coronavirus ; severe acute respiratory syndrome ; SARS ; SARS-CoV
刊名: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
发表日期: 2003-12-26
DOI: 10.1016/j.bbrc.2003.11.064
卷: 312, 期:4, 页:1290-1296
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemistry & Molecular Biology ; Biophysics
研究领域[WOS]: Biochemistry & Molecular Biology ; Biophysics
关键词[WOS]: ACUTE RESPIRATORY SYNDROME ; OUTBREAK ; PROTEIN
英文摘要:

An enhanced polymerase chain reaction (PCR) assay to detect the coronavirus associated with severe acute respiratory syndrome (SARS-CoV) was developed in which a target gene pre-amplification step preceded TaqMan real-time fluorescent PCR. Clinical samples were collected from 120 patients diagnosed as suspected or probable SARS cases and analyzed by conventional PCR followed by agarose gel electrophoresis, conventional TaqMan real-time PCR, and our enhanced TaqMan real-time PCR assays. An amplicon of the size expected from SARS-CoV was obtained from 28/120 samples using the enhanced real-time PCR method. Conventional PCR and real-time PCR alone identified fewer SARS-CoV positive cases. Results were confirmed by viral culture in 3/28 cases. The limit of detection of the enhanced real-time PCR method was 10(2)-fold higher than the standard real-time PCR assay and 10(7)-fold higher than conventional PCR methods. The increased sensitivity of the assay may help control the spread of the disease during future SARS outbreaks. (C) 2003 Elsevier Inc. All rights reserved.

语种: 英语
WOS记录号: WOS:000187252300064
Citation statistics:
内容类型: 期刊论文
版本: 出版稿
URI标识: http://ir.bjmu.edu.cn/handle/400002259/53670
Appears in Collections:基础医学院_神经生物学系_期刊论文

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作者单位: 1.Peking Univ, Neurosci Res Inst, Key Lab Neurosci, Beijing 100083, Peoples R China
2.Natl Emergency Act SARS Res, Beijing Grp, Beijing, Peoples R China
3.Hong Kong DNA Chips Ltd, Cosmos Ctr, Kowloon, Hong Kong, Peoples R China
4.Peking Univ, Hlth Sci Ctr, Dept Neurobiol, Beijing 100083, Peoples R China
5.Chinese Univ Hong Kong, Prince Wales Hosp, Dept Microbiol, Hong Kong, Hong Kong, Peoples R China

Recommended Citation:
Lau, LT,Fung, YWW,Wong, FPF,et al. A real-time PCR for SARS-coronavirus incorporating target gene pre-amplification[J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,2003,312(4):1290-1296.
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