A novel coronavirus associated with severe acute respiratory syndrome
A novel coronavirus associated with severe acute respiratory syndrome
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DOI：
10.1056/nejmoa030781
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发表时间：
2003-05-15
影响因子：
158.5
通讯作者：
Anderson, LJ
中科院分区：
文献类型：
--
作者：
Ksiazek, TG;Erdman, D;Anderson, LJ
背景：严重急性呼吸系统综合征（SARS）的全球暴发与来自中国广东省的一名患病卫生保健工作者的暴露有关。我们进行了研究以确定这次暴发的病原。方法我们收集了来自7个国家的患者的临床标本，并使用病毒分离技术、电子显微镜和组织学研究、分子和血清学分析对其进行检测，试图确定广泛的潜在病原体。结果所有呼吸道病原菌的鉴定结果均不一致。然而，从符合SARS病例定义的患者中分离出了一种新型冠状病毒。用咽拭子标本接种的Vero E6细胞具有细胞病理学特征。电镜检查显示冠状病毒的超微结构特征。免疫组织化学和免疫荧光染色显示与I型冠状病毒多克隆抗体有反应性。设计共识冠状病毒引物，通过逆转录聚合酶链反应（RT-PCR）扩增聚合酶基因片段，获得一个序列，清楚地确定该分离物是一种独特的冠状病毒，仅与先前测序的冠状病毒有远亲关系。通过特异性诊断RT-PCR引物，我们在来自多个地点的12名患者中鉴定出几个相同的核苷酸序列，这一发现与点源暴发一致。用新分离物进行的间接荧光抗体试验和酶联免疫吸附试验已被用于证明病毒特异性血清学反应。这种病毒以前可能从未在美国人群中传播过。结论一种新型冠状病毒与此次疫情有关，有证据表明该病毒在SARS发病中具有病原学作用。由于Carlo Urbani博士的去世，我们建议将我们的第一个分离株命名为sars相关冠状病毒的Urbani菌株。
BACKGROUNDA worldwide outbreak of severe acute respiratory syndrome (SARS) has been associated with exposures originating from a single ill health care worker from Guangdong Province, China. We conducted studies to identify the etiologic agent of this outbreak.METHODSWe received clinical specimens from patients in seven countries and tested them, using virus-isolation techniques, electron-microscopical and histologic studies, and molecular and serologic assays, in an attempt to identify a wide range of potential pathogens.RESULTSNone of the previously described respiratory pathogens were consistently identified. However, a novel coronavirus was isolated from patients who met the case definition of SARS. Cytopathological features were noted in Vero E6 cells inoculated with a throat-swab specimen. Electron-microscopical examination revealed ultrastructural features characteristic of coronaviruses. Immunohistochemical and immunofluorescence staining revealed reactivity with group I coronavirus polyclonal antibodies. Consensus coronavirus primers designed to amplify a fragment of the polymerase gene by reverse transcription-polymerase chain reaction (RT-PCR) were used to obtain a sequence that clearly identified the isolate as a unique coronavirus only distantly related to previously sequenced coronaviruses. With specific diagnostic RT-PCR primers we identified several identical nucleotide sequences in 12 patients from several locations, a finding consistent with a point-source outbreak. Indirect fluorescence antibody tests and enzyme-linked immunosorbent assays made with the new isolate have been used to demonstrate a virus-specific serologic response. This virus may never before have circulated in the U.S. population.CONCLUSIONSA novel coronavirus is associated with this outbreak, and the evidence indicates that this virus has an etiologic role in SARS. Because of the death of Dr. Carlo Urbani, we propose that our first isolate be named the Urbani strain of SARS-associated coronavirus.