Detecting SARS-CoV-2 BA.2,BA.4,and BA.5 Variants Utilizing a Robust RT-RPA-CRISPR/Cas12a-Based Method-China,2023

查看全文 作　　者：Meihui [1]Luo;Yang [2]Pan;Yaqing [3]He;Ruhan [1]A;Changcheng [1]Wu;Baoying [1]Huang;Roujian [1]Lu;Li [1]Zhao;Bo [3]Peng;Fei [1]Ye;Huijuan [1]Wang;Yuda [1,4]Chen;Zhen [1,5]Li;Daitao [2]Zhang;Wenling [1]Wang;Wenjie [1]Tan 机构地区：[1]MHC Key Laboratory of Biosafety,National Institute for Viral Disease Control and Prevention,China CDC,Beijing,China;[2]Beijing Center for Disease Prevention and Control,Beijing,China;[3]Microbiology Laboratory,Shenzhen Center for Disease Control and Prevention,Shenzhen City,Guangdong Province,China;[4]School of Public Health,Baotou Medical College,Baotou City,Inner Mongolia Autonomous Region,China;[5]Collaborative Innovation Centre for Regenerative Medicine and Medical BioResourse Development and Application Co-constructed by the Province and Ministry,Guangxi Medical University,Guangxi Zhuang Autonomous Region,China 出　　处：《China CDC weekly》索引2023年第5卷第26期,共8页 基　　金：Supported by the National Key Research and Development Program of China(2021YFC2300101,2021YFC0863300,2022YFC 2304101,2022YFC2303401). 摘　　要：Introduction:Since 2019,numerous variants of concern for severe acute respiratory syndrome virus 2(SARS-CoV-2)have emerged,leading to significant outbreaks.The development of novel,highly accurate,and rapid detection techniques for these new SARSCoV-2 variants remains a primary focus in the ongoing efforts to control and prevent the coronavirus disease 2019(COVID-19)pandemic.Methods:Reverse transcription-recombinase polymerase amplification combined with the clustered regularly interspaced short palindromic repeatsassociated protein 12a(CRISPR/Cas12a)system was used to validate the detection of the Omicron BA.2,BA.4,and BA.5 variants of SARS-CoV-2.Results:Our results demonstrate that the CRISPR/Cas12a assay is capable of effectively detecting the SARS-CoV-2 BA.2,BA.4,and BA.5 variants with a limit of detection of 10,1,and 10 copies/μL,respectively.Importantly,our assay successfully differentiated the three SARS-CoV-2 Omicron strains from one another.Additionally,we evaluated 46 SARS-CoV-2 positive clinical samples consisting of BA.2(n=20),BA.4(n=6),and BA.5(n=20)variants,and the sensitivity of our assay ranged from 90%to 100%,while the specificity was 100%.Discussion:This research presents a swift and reliable CRISPR-based method that may be employed to track the emergence of novel SARS-CoV-2 variants. 关 键 词：CRISPR/Cas LIMIT SPECIFICITY