Early diagnosis of SARS coronavirus infection by real time RT-PCR.
Early diagnosis of SARS coronavirus infection by real time RT-PCR.
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DOI：
10.1016/j.jcv.2003.08.004
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发表时间：
2003-12
期刊：
影响因子：
--
通讯作者：
Peiris JS
中科院分区：
文献类型：
--
作者：
Poon LL;Chan KH;Wong OK;Yam WC;Yuen KY;Guan Y;Lo YM;Peiris JS
背景：一种新型冠状病毒最近被确定为严重急性呼吸系统综合征（SARS）的病原。目前用于检测sars冠状病毒（SARS-Cov）的分子检测方法在疾病的早期阶段灵敏度较低。目的：通过优化病毒RNA提取方法和实时定量RT-PCR技术，建立并评价一种灵敏的SARS诊断试剂。研究设计：从经血清学证实感染SARS冠状病毒的SARS患者发病后第1-3天收集50份鼻咽吸入样本，并对30份阴性对照样本进行研究。对样品进行检测：(1)采用常规RNA提取方法进行第一代常规RT-PCR检测（Lancet 361(2003) 1319），(2)采用改进的RNA提取方法进行第一代常规RT-PCR检测，(3)采用改进的RNA提取方法进行实时定量RT-PCR检测。结果：在患病前3天收集的50份NPA标本中，11份（22%）在我们的第一代RT-PCR检测中呈阳性。对RNA提取方案进行修改后，22份（44%）样品在常规RT-PCR检测中呈阳性。将改进的RNA提取方法与实时荧光定量PCR技术相结合，实时荧光定量PCR检测阳性40例（80%）。阴性对照未见阳性信号。结论：优化RNA提取方法，应用实时定量RT-PCR技术，可大大提高检测对SARS早期诊断的敏感性。
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.
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DOI：
10.1016/s0140-6736(03)13077-2
发表时间：
2003-04-19
期刊：
Lancet (London, England)
影响因子：
--
作者：
Peiris JS;Lai ST;Poon LL;Guan Y;Yam LY;Lim W;Nicholls J;Yee WK;Yan WW;Cheung MT;Cheng VC;Chan KH;Tsang DN;Yung RW;Ng TK;Yuen KY;SARS study group
通讯作者：
SARS study group
DOI：
10.1016/s0140-6736(03)13967-0
发表时间：
2003-07-26
期刊：
Lancet (London, England)
影响因子：
--
作者：
Kuiken T;Fouchier RA;Schutten M;Rimmelzwaan GF;van Amerongen G;van Riel D;Laman JD;de Jong T;van Doornum G;Lim W;Ling AE;Chan PK;Tam JS;Zambon MC;Gopal R;Drosten C;van der Werf S;Escriou N;Manuguerra JC;Stöhr K;Peiris JS;Osterhaus AD
通讯作者：
Osterhaus AD
影响因子：
8.8
作者：
Kaiser, L;Briones, MS;Hayden, FG
通讯作者：
Hayden, FG
DOI：
10.1016/s0140-6736(03)13412-5
发表时间：
2003-05-24
期刊：
Lancet (London, England)
影响因子：
--
作者：
Peiris JS;Chu CM;Cheng VC;Chan KS;Hung IF;Poon LL;Law KI;Tang BS;Hon TY;Chan CS;Chan KH;Ng JS;Zheng BJ;Ng WL;Lai RW;Guan Y;Yuen KY;HKU/UCH SARS Study Group
通讯作者：
HKU/UCH SARS Study Group
影响因子：
158.5
作者：
Poutanen, SM;Low, DE;McGeer, AJ
通讯作者：
McGeer, AJ