SARS-CoV‑۲ and Its Omicron Variants Detection with RT-RPA -CRISPR/Cas۱۳a-Based Method at Room Temperature
- سال انتشار: 1402
- محل انتشار: مجله گزارش های بیوشیمی و زیست شناسی مولکولی، دوره: 12، شماره: 3
- کد COI اختصاصی: JR_RBMB-12-3_008
- زبان مقاله: انگلیسی
- تعداد مشاهده: 56
نویسندگان
The First Clinical College of Xiangnan University, the First People's Hospital of Chenzhou Affiliated Xiangnan University, Chenzhou ۴۲۳۰۰۰, Hunan, China & Translational Medicine Institute, the First People's Hospital of Chenzhou, Hengyang Medical School,
The First Clinical College of Xiangnan University, the First People's Hospital of Chenzhou Affiliated Xiangnan University, Chenzhou ۴۲۳۰۰۰, Hunan, China & The Oncology Department of the First People's Hospital of Chenzhou, Chenzhou ۴۲۳۰۰۰, Hunan, China.
The First Clinical College of Xiangnan University, the First People's Hospital of Chenzhou Affiliated Xiangnan University, Chenzhou ۴۲۳۰۰۰, Hunan, China & Translational Medicine Institute, the First People's Hospital of Chenzhou, Hengyang Medical School,
The First Clinical College of Xiangnan University, the First People's Hospital of Chenzhou Affiliated Xiangnan University, Chenzhou ۴۲۳۰۰۰, Hunan, China & Translational Medicine Institute, the First People's Hospital of Chenzhou, Hengyang Medical School,
The First Clinical College of Xiangnan University, the First People's Hospital of Chenzhou Affiliated Xiangnan University, Chenzhou ۴۲۳۰۰۰, Hunan, China & Translational Medicine Institute, the First People's Hospital of Chenzhou, Hengyang Medical School,
Translational Medicine Institute, the First People's Hospital of Chenzhou, Hengyang Medical School, University of South China, Chenzhou ۴۲۳۰۰۰, Hunan, China & National Engineering Research Center of Personalized Diagnostic and Therapeutic Technology, Huna
The First Clinical College of Xiangnan University, the First People's Hospital of Chenzhou Affiliated Xiangnan University, Chenzhou ۴۲۳۰۰۰, Hunan, China & Translational Medicine Institute, the First People's Hospital of Chenzhou, Hengyang Medical School,
چکیده
Background: The outbreak of severe acute respiratory syndrome coronavirus ۲ (SARS-CoV-۲) has triggered a global health crisis, with genetic mutations and evolution further creating uncertainty about epidemic risk. It is imperative to rapidly determine the nucleic acid sequence of SARS-CoV-۲ and its variants to combat the coronavirus pandemic. Our goal was to develop a rapid, room-temperature, point-of-care (POC) detection system to determine the nucleic acid sequences of SARS-CoV-۲ isolates, especially omicron variants. Methods: Based on the conserved nucleotide sequence of SARS-CoV-۲, bioinformatics software was used to analyze, design, and screen optimal enzymatic isothermal amplification primers and efficient CRISPR RNAs (crRNAs) of CRISPR/Cas۱۳a to the target sequences. Reverse transcription-recombinase polymerase amplification (RT-RPA) was used to amplify the virus, and CRISPR/Cas۱۳a-crRNA was used to cleave the SARS-CoV-۲ target sequence. The sensitivity of nucleic acid detection was assessed by serial dilution of plasmid templates. All reactions were performed at room temperature. Results: RT-RPA, combined with CRISPR/Cas۱۳a, can detect the SARS-CoV-۲ with a minimum content of ۱۰۲ copies/μL, and can effectively distinguish between the original strain and the Omicron variant with a minimum limit of detection (LOD) of ۱۰۳ copies/μL. Conclusion: The method developed in this study has potential application in clinical detection of SARS-CoV-۲ and its omicron variants.کلیدواژه ها
CRISPR/Cas۱۳a, Omicron variants, RT-RPA, SARS-CoV‑۲.اطلاعات بیشتر در مورد COI
COI مخفف عبارت CIVILICA Object Identifier به معنی شناسه سیویلیکا برای اسناد است. COI کدی است که مطابق محل انتشار، به مقالات کنفرانسها و ژورنالهای داخل کشور به هنگام نمایه سازی بر روی پایگاه استنادی سیویلیکا اختصاص می یابد.
کد COI به مفهوم کد ملی اسناد نمایه شده در سیویلیکا است و کدی یکتا و ثابت است و به همین دلیل همواره قابلیت استناد و پیگیری دارد.