Identification of a novel coronavirus in patients with severe acute respiratory syndrome
Identification of a novel coronavirus in patients with severe acute respiratory syndrome
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DOI：
10.1056/nejmoa030747
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发表时间：
2003-05-15
影响因子：
158.5
通讯作者：
Doerr, HW
中科院分区：
文献类型：
--
作者：
Drosten, C;G端nther, S;Doerr, HW
研究背景严重急性呼吸综合征（SARS）是一种新的临床疾病。方法应用细胞培养和分子生物学技术，从SARS患者的临床标本中寻找未知病毒，并在SARS患者中鉴定出一种新的冠状病毒。在细胞培养物中分离病毒，并通过基于聚合酶链式反应（PCR）的随机扩增程序获得300个核苷酸长度的序列。基因特征表明，该病毒与已知的冠状病毒只有远亲关系（50%至60%的核苷酸序列相同）。根据获得的序列，建立了特异性和敏感性检测新病毒的常规和实时荧光PCR方法。在SARS患者的各种临床标本中检测到病毒，但在对照组中未检测到。在痰液中发现了高达每毫升1亿个分子的高浓度病毒RNA。在急性期血浆和恢复期晚期粪便中也检测到极低浓度的病毒RNA。感染患者在分离病毒的Vero细胞上显示血清转化。结论新型冠状病毒可能在引起SARS中起作用。
BACKGROUNDThe severe acute respiratory syndrome (SARS) has recently been identified as a new clinical entity. SARS is thought to be caused by an unknown infectious agent.METHODSClinical specimens from patients with SARS were searched for unknown viruses with the use of cell cultures and molecular techniques.RESULTSA novel coronavirus was identified in patients with SARS. The virus was isolated in cell culture, and a sequence 300 nucleotides in length was obtained by a polymerase-chain-reaction (PCR)-based random-amplification procedure. Genetic characterization indicated that the virus is only distantly related to known coronaviruses (identical in 50 to 60 percent of the nucleotide sequence). On the basis of the obtained sequence, conventional and real-time PCR assays for specific and sensitive detection of the novel virus were established. Virus was detected in a variety of clinical specimens from patients with SARS but not in controls. High concentrations of viral RNA of up to 100 million molecules per milliliter were found in sputum. Viral RNA was also detected at extremely low concentrations in plasma during the acute phase and in feces during the late convalescent phase. Infected patients showed seroconversion on the Vero cells in which the virus was isolated.CONCLUSIONSThe novel coronavirus might have a role in causing SARS.