RNA提取
核糖核酸
2019年冠状病毒病(COVID-19)
严重急性呼吸综合征冠状病毒2型(SARS-CoV-2)
传染性
萃取(化学)
计算生物学
病毒学
溶解
核酸
病毒
临床诊断
生物
化学
计算机科学
基因
色谱法
医学
分子生物学
疾病
传染病(医学专业)
生物化学
病理
临床心理学
作者
Zhen Zhao,Haodong Cui,Wenxing Song,Xiaoling Ru,Wenhua Zhou,Xue-Feng Yu
标识
DOI:10.1101/2020.02.22.961268
摘要
1 Abstract The ongoing outbreak of the novel coronavirus disease 2019 (COVID-19) originating from Wuhan, China, draws worldwide concerns due to its long incubation period and strong infectivity. Although RT-PCR-based molecular diagnosis techniques are being widely applied for clinical diagnosis currently, timely and accurate diagnosis are still limited due to labour intensive and time-consuming operations of these techniques. To address the issue, herein we report the synthesis of poly (amino ester) with carboxyl groups (PC)-coated magnetic nanoparticles (pcMNPs), and the development of pcMNPs-based viral RNA extraction method for the sensitive detection of COVID-19 causing virus, the SARS-CoV-2. This method combines the lysis and binding steps into one step, and the pcMNPs-RNA complexes can be directly introduced into subsequent RT-PCR reactions. The simplified process can purify viral RNA from multiple samples within 20 min using a simple manual method or an automated high-throughput approach. By identifying two different regions ( ORFlab and N gene) of viral RNA, a 10-copy sensitivity and a strong linear correlation between 10 and 10 5 copies of SARS-CoV-2 pseudovirus particles are achieved. Benefitting from the simplicity and excellent performances, this new extraction method can dramatically reduce the turn-around time and operational requirements in current molecular diagnosis of COVID-19, in particular for the early clinical diagnosis.
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