Rapid Isothermal Amplification and Portable Detection System for SARS-CoV-2
Anurup Ganguli,Ariana Mostafa,Jacob Berger,Mehmet Y. Aydin,Fu Sun,Enrique Valera,Brian T. Cunningham,William P. King,Rashid Bashir +8 more
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TLDR
The details of an RT-LAMP isothermal assay for the detection of SARS-CoV-2 virus with performance comparable to currently approved tests using RT-PCR are provided, including the demonstration of a smartphone-based point-of-care device that can be used at the point of sample collection.Abstract:
The COVID-19 pandemic provides an urgent example where a gap exists between availability of state-of-the-art diagnostics and current needs. As assay details and primer sequences become widely known, many laboratories could perform diagnostic tests using methods such as RT-PCR or isothermal RT-LAMP amplification. A key advantage of RT-LAMP based approaches compared to RT-PCR is that RT-LAMP is known to be robust in detecting targets from unprocessed samples. In addition, RT-LAMP assays are performed at a constant temperature enabling speed, simplicity, and point-of-use testing. Here, we provide the details of an RT-LAMP isothermal assay for the detection of SARS-CoV-2 virus with performance comparable to currently approved tests using RT-PCR. We characterize the assay by introducing swabs in virus spiked synthetic nasal fluids, moving the swab to viral transport medium (VTM), and using a volume of that VTM for performing the amplification without an RNA extraction kit. The assay has a Limit-of-Detection (LOD) of 50 RNA copies/μL in the VTM solution within 20 minutes, and LOD of 5000 RNA copies/μL in the nasal solution. Additionally, we show the utility of this assay for real-time point-of-use testing by demonstrating detection of SARS-CoV-2 virus in less than 40 minutes using an additively manufactured cartridge and a smartphone-based reader. Finally, we explore the speed and cost advantages by comparing the required resources and workflows with RT-PCR. This work could accelerate the development and availability of SARS-CoV-2 diagnostics by proving alternatives to conventional laboratory benchtop tests.read more
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References
More filters
Journal ArticleDOI
Genomic characterisation and epidemiology of 2019 novel coronavirus: implications for virus origins and receptor binding.
Roujian Lu,Xiang Zhao,Juan Li,Peihua Niu,Bo Yang,Honglong Wu,Wenling Wang,Hao Song,Baoying Huang,Na Zhu,Yuhai Bi,Xuejun Ma,Faxian Zhan,Liang Wang,Tao Hu,Hong Zhou,Zhenhong Hu,Weimin Zhou,Li Zhao,Jing Chen,Yao Meng,Ji Wang,Yang Lin,Jianying Yuan,Zhihao Xie,Jinmin Ma,William J. Liu,Dayan Wang,Wenbo Xu,Edward C. Holmes,George F. Gao,George F. Gao,Guizhen Wu,Weijun Chen,Weifeng Shi,Wenjie Tan,Wenjie Tan +36 more
TL;DR: The phylogenetic analysis suggests that bats might be the original host of this virus, an animal sold at the seafood market in Wuhan might represent an intermediate host facilitating the emergence of the virus in humans.
Journal ArticleDOI
Loop-mediated isothermal amplification of DNA
Tsugunori Notomi,Hiroto Okayama,Harumi Otawara-shi Masubuchi,Toshihiro Yonekawa,Keiko Watanabe,Nobuyuki Amino,Tetsu Hase +6 more
TL;DR: A novel method that amplifies DNA with high specificity, efficiency and rapidity under isothermal conditions that employs a DNA polymerase and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA.
Journal ArticleDOI
SARS-CoV-2 Viral Load in Upper Respiratory Specimens of Infected Patients.
Lirong Zou,Feng Ruan,Mingxing Huang,Lijun Liang,Huitao Huang,Zhongsi Hong,Jianxiang Yu,Min Kang,Yingchao Song,Jinyu Xia,Qianfang Guo,Tie Song,Jianfeng He,Hui-Ling Yen,Malik Peiris,Jie Wu +15 more
TL;DR: Results of an analysis of nasal and throat swabs from 17 patients in Zhuhai, China, who had received a diagnosis of Covid-19 and found SARS-CoV-2 Viral Load in Upper Respiratory Specimens positive.
Journal ArticleDOI
CRISPR-Cas12-based detection of SARS-CoV-2.
James Paul Broughton,Xianding Deng,Guixia Yu,Clare L Fasching,Venice Servellita,Jasmeet Singh,Xin Miao,Jessica Streithorst,Andrea Granados,Alicia Sotomayor-Gonzalez,Kelsey C. Zorn,Allan Gopez,Elaine Hsu,Wei Gu,Steve Miller,Chao Yang Pan,Hugo Guevara,Debra A. Wadford,Janice S. Chen,Charles Y. Chiu +19 more
TL;DR: The CRISPR-based DETECTR assay provides a visual and faster alternative to the US Centers for Disease Control and Prevention SARS-CoV-2 real-time RT–PCR assay, with 95% positive predictive agreement and 100% negative predictive agreement.
Journal ArticleDOI
Field-deployable viral diagnostics using CRISPR-Cas13
Cameron Myhrvold,Cameron Myhrvold,Catherine A. Freije,Catherine A. Freije,Jonathan S. Gootenberg,Omar O. Abudayyeh,Hayden C. Metsky,Ann Durbin,Ann Durbin,Max J. Kellner,Amanda L Tan,Lauren M. Paul,Leda Parham,Kimberly García,Kayla G. Barnes,Kayla G. Barnes,Bridget Chak,Bridget Chak,Adriano Mondini,Maurício Lacerda Nogueira,Sharon Isern,Scott F. Michael,Ivette Lorenzana,Nathan L. Yozwiak,Nathan L. Yozwiak,Bronwyn MacInnis,Bronwyn MacInnis,Irene Bosch,Irene Bosch,Lee Gehrke,Lee Gehrke,Feng Zhang,Pardis C. Sabeti +32 more
TL;DR: The Cas13-based SHERLOCK platform can detect Zika virus and dengue virus in patient samples at concentrations as low as 1 copy per microliter and can distinguish the four DENV serotypes, as well as region-specific strains of ZIKV from the 2015–2016 pandemic.
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