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Rapid Detection of SARS-CoV-2 Virus Using Dual Reverse Transcriptional Colorimetric Loop-Mediated Isothermal Amplification.
Ji, Chao; Xue, Shuxia; Yu, Min; Liu, Jinyu; Zhang, Qin; Zuo, Feng; Zheng, Qiuyue; Zhao, Liangjuan; Zhang, Hongwei; Cao, Jijuan; Wang, Ke; Liu, Wei; Zheng, Wenjie.
  • Ji C; State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan Agricultural University, Kunming 650201, P. R. China.
  • Xue S; Key Laboratory for Agro-Biodiversity and Pest Control of Ministry of Education, Yunnan Agricultural University, Kunming 650201, P. R. China.
  • Yu M; Laboratory for Quality Control and Traceability of Food, Tianjin Normal University, Tianjin 300387, P. R. China.
  • Liu J; Laboratory for Quality Control and Traceability of Food, Tianjin Normal University, Tianjin 300387, P. R. China.
  • Zhang Q; Department of Gynecologic Oncology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin 300060, P. R. China.
  • Zuo F; Laboratory for Quality Control and Traceability of Food, Tianjin Normal University, Tianjin 300387, P. R. China.
  • Zheng Q; Laboratory for Quality Control and Traceability of Food, Tianjin Normal University, Tianjin 300387, P. R. China.
  • Zhao L; Tianjin Customs District, Tianjin 300308, China.
  • Zhang H; Key Laboratory of Biotechnology and Bioresources Utilization of Ministry of Education, College of Life Science, Dalian Minzu University, Dalian 116600, P. R. China.
  • Cao J; Tianjin Customs District, Tianjin 300308, China.
  • Wang K; Tianjin Customs District, Tianjin 300308, China.
  • Liu W; Key Laboratory of Biotechnology and Bioresources Utilization of Ministry of Education, College of Life Science, Dalian Minzu University, Dalian 116600, P. R. China.
  • Zheng W; Department of Gynecologic Oncology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin 300060, P. R. China.
ACS Omega ; 6(13): 8837-8849, 2021 Apr 06.
Article in English | MEDLINE | ID: covidwho-1177080
ABSTRACT
The outbreak and pandemic of COVID-19, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has developed into a public health emergency of international concern. The rapid and accurate detection of the virus is a critical means to prevent and control the disease. Herein, we provide a novel, rapid, and simple approach, named dual reverse transcriptional colorimetric loop-mediated isothermal amplification (dRT-cLAMP) assay, to accelerate the detection of the SARS-CoV-2 virus without using expensive equipment. The result of this assay is shown by color change and is easily detected by the naked eye. To improve the detection accuracy, we included two primer sets that specifically target the viral orf1ab and N genes in the same reaction mixture. Our assay can detect the synthesized SARS-CoV-2 N and orf1ab genes at a low level of 100 copies/µL. Sequence alignment analysis of the two synthesized genes and those of 9968 published SARS-CoV-2 genomes and 17 genomes of other pathogens from the same infection site or similar symptoms as COVID-19 revealed that the primers for the dRT-cLAMP assay are highly specific. Our assay of 27 clinical samples of SARS-CoV-2 virus and 27 standard-added environmental simulation samples demonstrated that compared to the commercial kits, the consistency of the positive, negative, and probable clinical samples was 100, 92.31, and 44.44%, respectively. Moreover, our results showed that the positive, but not negative, standard-added samples displayed a naked-eye-detectable color change. Together, our results demonstrate that the dRT-cLAMP assay is a feasible detection assay for SARS-CoV-2 virus and is of great significance since rapid onsite detection of the virus is urgently needed at the ports of entry, health care centers, and for internationally traded goods.

Full text: Available Collection: International databases Database: MEDLINE Type of study: Prognostic study Language: English Journal: ACS Omega Year: 2021 Document Type: Article

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Full text: Available Collection: International databases Database: MEDLINE Type of study: Prognostic study Language: English Journal: ACS Omega Year: 2021 Document Type: Article