RNA提取
经济短缺
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)
核糖核酸
人口
医学
萃取(化学)
病毒学
生物
色谱法
化学
病理
疾病
传染病(医学专业)
环境卫生
生物化学
哲学
政府(语言学)
基因
语言学
作者
Adolfo Marcelo,Fiorella Coronado-Marquina,Jairo Andrés Méndez Rico,Maria C.B. Mendoza,Nancy Rojas‐Serrano,Paulo Vitor Marques Simas,César Sánchez,Jan Felix Drexler
出处
期刊:PLOS ONE
[Public Library of Science]
日期:2021-03-24
卷期号:16 (3): e0248885-e0248885
被引量:27
标识
DOI:10.1371/journal.pone.0248885
摘要
One of the biggest challenges during the pandemic has been obtaining and maintaining critical material to conduct the increasing demand for molecular tests. Sometimes, the lack of suppliers and the global shortage of these reagents, a consequence of the high demand, make it difficult to detect and diagnose patients with suspected SARS-CoV-2 infection, negatively impacting the control of virus spread. Many alternatives have enabled the continuous processing of samples and have presented a decrease in time and cost. These measures thus allow broad testing of the population and should be ideal for controlling the disease. In this sense, we compared the SARS-CoV-2 molecular detection effectiveness by Real time RT-PCR using two different protocols for RNA extraction. The experiments were conducted in the National Institute of Health (INS) from Peru. We compared Ct values average (experimental triplicate) results from two different targets, a viral and internal control. All samples were extracted in parallel using a commercial kit and our alternative protocol–samples submitted to proteinase K treatment (3 μg/μL, 56°C for 10 minutes) followed by thermal shock (98°C for 5 minutes followed by 4°C for 2 minutes); the agreement between results was 100% in the samples tested. In addition, we compared the COVID-19 positivity between six epidemiological weeks: the initial two in that the Real time RT-PCR reactions were conducted using RNA extracted by commercial kit, followed by two other using RNA obtained by our kit-free method, and the last two using kit once again; they did not differ significantly. We concluded that our in-house method is an easy, fast, and cost-effective alternative method for extracting RNA and conducing molecular diagnosis of COVID-19.
科研通智能强力驱动
Strongly Powered by AbleSci AI