iSCAN: An RT-LAMP-coupled CRISPR-Cas12 module for rapid, sensitive detection of SARS-CoV-2.
Name:
iscan_1-s2.0-S0168170220310364-main.pdf
Size:
2.345Mb
Format:
PDF
Description:
Accepted Article
Type
ArticleAuthors
Ali, Zahir
Aman, Rashid

Mahas, Ahmed

Rao, Gundra Sivakrishna
Tehseen, Muhammad

Marsic, Tin
Salunke, Rahul Pandurang
Subudhi, Amit K
Hala, Sharif M
Hamdan, Samir

Pain, Arnab

Alofi, Fadwa S
Alsomali, Afrah
Hashem, Anwar M
Khogeer, Asim
Almontashiri, Naif A M
Abedalthagafi, Malak
Hassan, Norhan
Mahfouz, Magdy M.

KAUST Department
Biological and Environmental Science and Engineering (BESE) DivisionBioscience Program
Center for Desert Agriculture
Laboratory for Genome Engineering
Laboratory for Genome Engineering and Synthetic Biology, Division of Biological Sciences, 4700 King Abdullah University of Science and Technology, Thuwal, 23955-6900, Saudi Arabia.
Laboratory of DNA Replication and Recombination
Pathogen Genomics Laboratory
Pathogen Genomics Laboratory, BESE Division, King Abdullah University of Science and Technology (KAUST), Thuwal, Saudi Arabia.
Plant Science
Date
2020-08-18Online Publication Date
2020-08-18Print Publication Date
2020-10Embargo End Date
2021-08-22Submitted Date
2020-07-04Permanent link to this record
http://hdl.handle.net/10754/664788
Metadata
Show full item recordSummary
This record has been merged with an existing record at: http://hdl.handle.net/10754/663651.
Abstract
The COVID-19 pandemic caused by SARS-CoV-2 affects all aspects of human life. Detection platforms that are efficient, rapid, accurate, specific, sensitive, and user friendly are urgently needed to manage and control the spread of SARS-CoV-2. RT-qPCR based methods are the gold standard for SARS-CoV-2 detection. However, these methods require trained personnel, sophisticated infrastructure, and a long turnaround time, thereby limiting their usefulness. Reverse transcription-loop-mediated isothermal amplification (RT-LAMP), a one-step nucleic acid amplification method conducted at a single temperature, has been used for colorimetric virus detection. CRISPR-Cas12 and CRISPR-Cas13 systems, which possess collateral activity against ssDNA and RNA, respectively, have also been harnessed for virus detection. Here, we built an efficient, rapid, specific, sensitive, user-friendly SARS-CoV-2 detection module that combines the robust virus amplification of RT-LAMP with the specific detection ability of SARS-CoV-2 by CRISPR-Cas12. Furthermore, we combined the RT-LAMP-CRISPR-Cas12 module with lateral flow cells to enable highly efficient point-of-care SARS-CoV-2 detection. Our iSCAN SARS-CoV-2 detection module, which exhibits the critical features of a robust molecular diagnostic device, should facilitate the effective management and control of COVID-19.Citation
Ali, Z., Aman, R., Mahas, A., Rao, G. S., Tehseen, M., Marsic, T., … Mahfouz, M. M. (2020). iSCAN: An RT-LAMP-coupled CRISPR-Cas12 module for rapid, sensitive detection of SARS-CoV-2. Virus Research, 198129. doi:10.1016/j.virusres.2020.198129Sponsors
We would like to thank members of the genome engineering and synthetic biology laboratory for insightful discussions and technical support. We thank Professor Andrew Ellington for providing the RTx and BstI clones. This work was supported, in part, by the Smart Health Initiative at KAUST and the IAF grant from the KAUST IED to MM. The samples were collected from patients providing verbal consent to the treating doctor after they read the patient consent form, or the doctor explained the content of consent form to the patient. Hence, as per the approved protocol, the doctors took samples only after receiving verbal consent from a concerned patient (due to infection hazard and no family member being available next to the patients due to infection control guidelines) and the treating doctor signed on the consent form on behalf of the patient before any sample collection took place. We have obtained the necessary ethical approval from the Saudi-MOH and the Mass Gathering directorate in Riyadh, Saudi Arabia, for the collection of the samples (Saudi MOH Approval #: H-02-K-076-0420-285, dated 13.04.2020; KAUST Approval #: 20IBEC14). We thank Sara Mfarrej and Fathia Ben Rached for their help during the handling of the clinical samples in KAUST.Publisher
Elsevier BVJournal
Virus researchPubMed ID
32822689Additional Links
https://linkinghub.elsevier.com/retrieve/pii/S0168170220310364ae974a485f413a2113503eed53cd6c53
10.1016/j.virusres.2020.198129
Scopus Count
Related articles
- Loop mediated isothermal amplification (LAMP) assays as a rapid diagnostic for COVID-19.
- Authors: Kashir J, Yaqinuddin A
- Issue date: 2020 Aug
- Contamination-free visual detection of SARS-CoV-2 with CRISPR/Cas12a: A promising method in the point-of-care detection.
- Authors: Chen Y, Shi Y, Chen Y, Yang Z, Wu H, Zhou Z, Li J, Ping J, He L, Shen H, Chen Z, Wu J, Yu Y, Zhang Y, Chen H
- Issue date: 2020 Dec 1
- Development and Clinical Application of a Rapid and Sensitive Loop-Mediated Isothermal Amplification Test for SARS-CoV-2 Infection.
- Authors: Hu X, Deng Q, Li J, Chen J, Wang Z, Zhang X, Fang Z, Li H, Zhao Y, Yu P, Li W, Wang X, Li S, Zhang L, Hou T
- Issue date: 2020 Aug 26
- SARS-CoV-2 RNA Extraction Using Magnetic Beads for Rapid Large-Scale Testing by RT-qPCR and RT-LAMP.
- Authors: Klein S, Müller TG, Khalid D, Sonntag-Buck V, Heuser AM, Glass B, Meurer M, Morales I, Schillak A, Freistaedter A, Ambiel I, Winter SL, Zimmermann L, Naumoska T, Bubeck F, Kirrmaier D, Ullrich S, Barreto Miranda I, Anders S, Grimm D, Schnitzler P, Knop M, Kräusslich HG, Dao Thi VL, Börner K, Chlanda P
- Issue date: 2020 Aug 7
- A colorimetric RT-LAMP assay and LAMP-sequencing for detecting SARS-CoV-2 RNA in clinical samples.
- Authors: Dao Thi VL, Herbst K, Boerner K, Meurer M, Kremer LP, Kirrmaier D, Freistaedter A, Papagiannidis D, Galmozzi C, Stanifer ML, Boulant S, Klein S, Chlanda P, Khalid D, Barreto Miranda I, Schnitzler P, Kräusslich HG, Knop M, Anders S
- Issue date: 2020 Aug 12