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Detection of severe acute respiratory syndrome coronavirus 2 and influenza viruses based on CRISPR-Cas12a
Experimental Biology and Medicine ( IF 2.8 ) Pub Date : 2020-11-05 , DOI: 10.1177/1535370220963793
Oraphan Mayuramart 1 , Pattaraporn Nimsamer 1 , Somruthai Rattanaburi 1 , Naphat Chantaravisoot 1 , Kritsada Khongnomnan 2 , Jira Chansaenroj 3 , Jiratchaya Puenpa 3 , Nungruthai Suntronwong 3 , Preeyaporn Vichaiwattana 3 , Yong Poovorawan 3 , Sunchai Payungporn 1, 4
Affiliation  

Due to the common symptoms of COVID-19, patients are similar to influenza-like illness. Therefore, the detection method would be crucial to discriminate between SARS-CoV-2 and influenza virus-infected patients. In this study, CRISPR-Cas12a-based detection was applied for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A virus, and influenza B virus which would be a practical and attractive application for screening of patients with COVID-19 and influenza in areas with limited resources. The limit of detection for SARS-CoV-2, influenza A, and influenza B detection was 10, 103, and 103 copies/reaction, respectively. Moreover, the assays yielded no cross-reactivity against other respiratory viruses. The results revealed that the detection of influenza virus and SARS-CoV-2 by using RT-RPA and CRISPR-Cas12a technology reaches 96.23% sensitivity and 100% specificity for SARS-CoV-2 detection. The sensitivity for influenza virus A and B detections was 85.07% and 94.87%, respectively. In addition, the specificity for influenza virus A and B detections was approximately 96%. In conclusion, the RT-RPA with CRISPR-Cas12a assay was an effective method for the screening of influenza viruses and SARS-CoV-2 which could be applied to detect other infectious diseases in the future.



中文翻译:

基于CRISPR-Cas12a检测严重急性呼吸综合征冠状病毒2型和流感病毒

由于 COVID-19 的常见症状,患者类似于流感样疾病。因此,检测方法对于区分 SARS-CoV-2 和流感病毒感染患者至关重要。在这项研究中,基于 CRISPR-Cas12a 的检测被应用于检测严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2)、甲型流感病毒和乙型流感病毒,这将是筛查患有严重急性呼吸系统综合症的患者的实用且有吸引力的应用。资源有限地区的 COVID-19 和流感。SARS-CoV-2、甲型流感和乙型流感的检测限分别为 10、10 3和 10 3分别复制/反应。此外,这些检测对其他呼吸道病毒没有产生交叉反应。结果表明,使用RT-RPA和CRISPR-Cas12a技术检测流感病毒和SARS-CoV-2对SARS-CoV-2检测的敏感性达到96.23%,特异性达到100%。流感病毒 A 和 B 检测的灵敏度分别为 85.07% 和 94.87%。此外,流感病毒 A 和 B 检测的特异性约为 96%。总之,带有CRISPR-Cas12a检测的RT-RPA是筛选流感病毒和SARS-CoV-2的有效方法,未来可用于检测其他传染病。

更新日期:2020-11-06
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