Early diagnosis of SARS Coronavirus infection by real time RT-PCR

被引:147
作者
Poon, LLM [1 ]
Chan, KH
Wong, OK
Yam, WC
Yuen, KY
Guan, Y
Lo, YMD
Peiris, JSM
机构
[1] Univ Hong Kong, Queen Mary Hosp, Dept Microbiol, Hong Kong, Hong Kong, Peoples R China
[2] Queen Marys Hosp, Dept Microbiol, Hong Kong, Hong Kong, Peoples R China
[3] Chinese Univ Hong Kong, Dept Chem Pathol, Sha Tin, Hong Kong, Peoples R China
关键词
early diagnosis; SARS Coronavirus; real time RT-PCR;
D O I
10.1016/j.jcv.2003.08.004
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Background: A novel coronavirus was recently identified as the aetiological agent of Severe Acute Respiratory Syndrome (SARS). Molecular assays currently available for detection of SARS-coronavirus (SARS-Cov) have low sensitivity during the early stage of the illness. Objective: To develop and evaluate a sensitive diagnostic test for SARS by optimizing the viral RNA extraction methods and by applying real-time quantitative RT-PCR technology. Study design: 50 nasopharyngeal aspirate (NPA) samples collected from days 1-3 of disease onset from SARS patients in whom SARS CoV infections was subsequently serologically confirmed and 30 negative control samples were studied. Samples were tested by: (1) our first generation conventional RT-PCR assay with a routine RNA extraction method (Lancet 361 (2003) 1319), (2) our first generation conventional RT-PCR assay with a modified RNA extraction method, (3) a real-time quantitative RT-PCR assay with a modified RNA extraction method. Results: Of 50 NPA specimens collected during the first 3 days of illness, 11 (22%) were positive in our first generation RT-PCR assay. With a modification in the RNA extraction protocol, 22 (44%) samples were positive in the conventional RT-PCR assay. By combining the modified RNA extraction method and real-time quantitative PCR technology, 40 (80%) of these samples were positive in the real-time RT-PCR assay. No positive signal was observed in the negative controls. Conclusion: By optimizing RNA extraction methods and applying quantitative real time RT-PCR technologies, the sensitivity of tests for early diagnosis of SARS can be greatly enhanced. (C) 2003 Published by Elsevier B.V.
引用
收藏
页码:233 / 238
页数:6
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