Molecular Diagnosis of a Novel Coronavirus (2019-nCoV) Causing an Outbreak of Pneumonia.

引起肺炎暴发的新型冠状病毒 (2019-nCoV) 的分子诊断。

  • 影响因子:3.34
  • DOI:10.1093/clinchem/hvaa029
  • 作者列表:"Chu DKW","Pan Y","Cheng SMS","Hui KPY","Krishnan P","Liu Y","Ng DYM","Wan CKC","Yang P","Wang Q","Peiris M","Poon LLM
  • 发表时间:2020-04-01

BACKGROUND:A novel coronavirus of zoonotic origin (2019-nCoV) has recently been identified in patients with acute respiratory disease. This virus is genetically similar to SARS coronavirus and bat SARS-like coronaviruses. The outbreak was initially detected in Wuhan, a major city of China, but has subsequently been detected in other provinces of China. Travel-associated cases have also been reported in a few other countries. Outbreaks in health care workers indicate human-to-human transmission. Molecular tests for rapid detection of this virus are urgently needed for early identification of infected patients. METHODS:We developed two 1-step quantitative real-time reverse-transcription PCR assays to detect two different regions (ORF1b and N) of the viral genome. The primer and probe sets were designed to react with this novel coronavirus and its closely related viruses, such as SARS coronavirus. These assays were evaluated using a panel of positive and negative controls. In addition, respiratory specimens from two 2019-nCoV-infected patients were tested. RESULTS:Using RNA extracted from cells infected by SARS coronavirus as a positive control, these assays were shown to have a dynamic range of at least seven orders of magnitude (2x10-4-2000 TCID50/reaction). Using DNA plasmids as positive standards, the detection limits of these assays were found to be below 10 copies per reaction. All negative control samples were negative in the assays. Samples from two 2019-nCoV-infected patients were positive in the tests. CONCLUSIONS:The established assays can achieve a rapid detection of 2019n-CoV in human samples, thereby allowing early identification of patients.


背景: 最近在急性呼吸系统疾病患者中发现了人畜共患病起源的新型冠状病毒 (2019-nCoV)。这种病毒遗传类似于传染性非典型肺炎冠状病毒病毒和蝙蝠传染性非典型肺炎样冠状病毒病毒es.疫情最初在中国的一个主要城市武汉被发现,但随后在中国的其他省份被发现。其他几个国家也报告了与旅行相关的病例。卫生保健工作者的爆发表明人传人。为了早期识别感染患者,迫切需要快速检测这种病毒的分子检测。 方法: 我们开发了两个 1 步定量实时逆转录PCR检测方法来检测病毒基因组的两个不同区域 (ORF1b和N)。引物和探针组的目的是与这个新型冠状病毒及其密切相关的病毒,如传染性非典型肺炎冠状病毒.使用一组阳性和阴性对照评价这些试验。此外,对 2 例 2019-nCoV感染患者的呼吸道标本进行了检测。 结果: 传染性非典型肺炎冠状病毒感染的细胞中提取的RNA作为阳性对照,这些检测显示至少有 7 个数量级的动态范围 (2x10-4-2000 TCID50/反应)。使用DNA质粒作为阳性标准品,发现这些试验的检测限低于每个反应 10 个拷贝。所有阴性对照样品在试验中均为阴性。两名 2019 nCoV感染患者的样本在检测中呈阳性。 结论: 建立的检测方法可以实现人类样本中 2019n-CoV的快速检测,从而可以早期识别患者。



作者列表:["Lim J","Jeon S","Shin HY","Kim MJ","Seong YM","Lee WJ","Choe KW","Kang YM","Lee B","Park SJ"]

METHODS::Since mid-December of 2019, coronavirus disease 2019 (COVID-19) infection has been spreading from Wuhan, China. The confirmed COVID-19 patients in South Korea are those who came from or visited China. As secondary transmissions have occurred and the speed of transmission is accelerating, there are rising concerns about community infections. The 54-year old male is the third patient diagnosed with COVID-19 infection in Korea. He is a worker for a clothing business and had mild respiratory symptoms and intermittent fever in the beginning of hospitalization, and pneumonia symptoms on chest computerized tomography scan on day 6 of admission. This patient caused one case of secondary transmission and three cases of tertiary transmission. Hereby, we report the clinical findings of the index patient who was the first to cause tertiary transmission outside China. Interestingly, after lopinavir/ritonavir (Kaletra, AbbVie) was administered, β-coronavirus viral loads significantly decreased and no or little coronavirus titers were observed.

作者列表:["Zhang W","Du RH","Li B","Zheng XS","Yang XL","Hu B","Wang YY","Xiao GF","Yan B","Shi ZL","Zhou P"]

METHODS::In December 2019, a novel coronavirus (2019-nCoV) caused an outbreak in Wuhan, China, and soon spread to other parts of the world. It was believed that 2019-nCoV was transmitted through respiratory tract and then induced pneumonia, thus molecular diagnosis based on oral swabs was used for confirmation of this disease. Likewise, patient will be released upon two times of negative detection from oral swabs. However, many coronaviruses can also be transmitted through oral-fecal route by infecting intestines. Whether 2019-nCoV infected patients also carry virus in other organs like intestine need to be tested. We conducted investigation on patients in a local hospital who were infected with this virus. We found the presence of 2019-nCoV in anal swabs and blood as well, and more anal swab positives than oral swab positives in a later stage of infection, suggesting shedding and thereby transmitted through oral-fecal route. We also showed serology test can improve detection positive rate thus should be used in future epidemiology. Our report provides a cautionary warning that 2019-nCoV may be shed through multiple routes.

翻译标题与摘要 下载文献
作者列表:["Cheng ZJ","Shan J"]

METHODS::There is a current worldwide outbreak of a new type of coronavirus (2019-nCoV), which originated from Wuhan in China and has now spread to 17 other countries. Governments are under increased pressure to stop the outbreak spiraling into a global health emergency. At this stage, preparedness, transparency, and sharing of information are crucial to risk assessments and beginning outbreak control activities. This information should include reports from outbreak sites and from laboratories supporting the investigation. This paper aggregates and consolidates the virology, epidemiology, clinical management strategies from both English and Chinese literature, official news channels, and other official government documents. In addition, by fitting the number of infections with a single-term exponential model, we report that the infection is spreading at an exponential rate, with a doubling period of 1.8 days.