Rapid point-of-care detection of SARS-CoV-2 using reverse transcription loop-mediated isothermal amplification (RT-LAMP)

被引:102
作者
Mautner, Lena [1 ]
Baillie, Christin-Kirsty [1 ]
Herold, Heike Marie [1 ]
Volkwein, Wolfram [1 ]
Guertler, Patrick [1 ]
Eberle, Ute [1 ]
Ackermann, Nikolaus [1 ]
Sing, Andreas [1 ]
Pavlovic, Melanie [1 ]
Goerlich, Ottmar [1 ]
Busch, Ulrich [1 ]
Wassill, Lars [1 ]
Huber, Ingrid [1 ]
Baiker, Armin [1 ]
机构
[1] Bavarian Hlth & Food Safety Author, Vet Str 2, D-85764 Oberschleissheim, Germany
关键词
RT-LAMP; Point-of-care testing; SARS-CoV-2; COVID-19; Rapid testing; No RNA extraction; ORF8; Gene N;
D O I
10.1186/s12985-020-01435-6
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Background Fast, reliable and easy to handle methods are required to facilitate urgently needed point-of-care testing (POCT) in the current coronavirus pandemic. Life-threatening severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has rapidly spread all over the world, infecting more than 33,500,000 people and killing over 1 million of them as of October 2020. Infected individuals without any symptoms might still transfer the virus to others underlining the extraordinary transmissibility of this new coronavirus. In order to identify early infections effectively, treat patients on time and control disease spreading, rapid, accurate and onsite testing methods are urgently required. Results Here we report the development of a loop-mediated isothermal amplification (LAMP) based method to detect SARS-CoV-2 genes ORF8 and N directly from pharyngeal swab samples. The established reverse transcription LAMP (RT-LAMP) assay detects SARS-CoV-2 directly from pharyngeal swab samples without previous time-consuming and laborious RNA extraction. The assay is sensitive and highly specific for SARS-CoV-2 detection, showing no cross reactivity when tested on 20 other respiratory pathogens. The assay is 12 times faster and 10 times cheaper than routine reverse transcription real-time polymerase chain reaction, depending on the assay used. Conclusion The fast and easy to handle RT-LAMP assay amplifying specifically the genomic regions ORF8 and N of SARS-CoV-2 is ideally suited for POCT at e.g. railway stations, airports or hospitals. Given the current pandemic situation, rapid, cost efficient and onsite methods like the here presented RT-LAMP assay are urgently needed to contain the viral spread.
引用
收藏
页数:14
相关论文
共 22 条
[1]   Development of a reverse transcription-loop-mediated isothermal amplification as a rapid early-detection method for novel SARS-CoV-2 [J].
Baek, Yun Hee ;
Um, Jihye ;
Antigua, Khristine Joy C. ;
Park, Ji-Hyun ;
Kim, Yeonjae ;
Oh, Sol ;
Kim, Young-il ;
Choi, Won-Suk ;
Kim, Seong Gyu ;
Jeong, Ju Hwan ;
Chin, Bum Sik ;
Nicolas, Halcyon Dawn G. ;
Ahn, Ji-Young ;
Shin, Kyeong Seob ;
Choi, Young Ki ;
Park, Jun-Sun ;
Song, Min-Suk .
EMERGING MICROBES & INFECTIONS, 2020, 9 (01) :998-1007
[2]   Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR (Publication with Expression of Concern) [J].
Corman, Victor M. ;
Landt, Olfert ;
Kaiser, Marco ;
Molenkamp, Richard ;
Meijer, Adam ;
Chu, Daniel K. W. ;
Bleicker, Tobias ;
Bruenink, Sebastian ;
Schneider, Julia ;
Schmidt, Marie Luisa ;
Mulders, Daphne G. J. C. ;
Haagmans, Bart L. ;
van der Veer, Bas ;
van den Brink, Sharon ;
Wijsman, Lisa ;
Goderski, Gabriel ;
Romette, Jean-Louis ;
Ellis, Joanna ;
Zambon, Maria ;
Peiris, Malik ;
Goossens, Herman ;
Reusken, Chantal ;
Koopmans, Marion P. G. ;
Drosten, Christian .
EUROSURVEILLANCE, 2020, 25 (03) :23-30
[3]   A multiplexed RT-LAMP assay for detection of group M HIV-1 in plasma or whole blood [J].
Curtis, Kelly A. ;
Morrison, Daphne ;
Rudolph, Donna L. ;
Shankar, Anupama ;
Bloomfield, Laura S. P. ;
Switzer, William M. ;
Owen, S. Michele .
JOURNAL OF VIROLOGICAL METHODS, 2018, 255 :91-97
[4]   Theory for RNA Folding, Stretching, and Melting Including Loops and Salt [J].
Einert, Thomas R. ;
Netz, Roland R. .
BIOPHYSICAL JOURNAL, 2011, 100 (11) :2745-2753
[5]   Rapid detection of epidermal growth factor receptor mutations in lung cancer by the SMart-Amplification process [J].
Hoshi, Kanako ;
Takakura, Hideki ;
Mitani, Yasumasa ;
Tatsumi, Kenji ;
Momiyama, Nobuyoshi ;
Ichikawa, Yasushi ;
Togo, Shinji ;
Miyag, Toru ;
Kawai, Yuki ;
Kogo, Yasushi ;
Kikuch, Takeshi ;
Kato, Chiaki ;
Arakawa, Takahiro ;
Uno, Syuji ;
Cizdziel, Paul E. ;
Lezhava, Alexander ;
Ogawa, Noburou ;
Hayashizaki, Yoshihicle ;
Shimada, Hiroshi .
CLINICAL CANCER RESEARCH, 2007, 13 (17) :4974-4983
[6]   RT-LAMP for rapid diagnosis of coronavirus SARS-CoV-2 [J].
Huang, Wei E. ;
Lim, Boon ;
Hsu, Chia-Chen ;
Xiong, Dan ;
Wu, Wei ;
Yu, Yejiong ;
Jia, Huidong ;
Wang, Yun ;
Zeng, Yida ;
Ji, Mengmeng ;
Chang, Hong ;
Zhang, Xiuming ;
Wang, Hui ;
Cui, Zhanfeng .
MICROBIAL BIOTECHNOLOGY, 2020, 13 (04) :950-961
[7]   Rapid establishment of laboratory diagnostics for the novel coronavirus SARS-CoV-2 in Bavaria, Germany, February 2020 [J].
Konrad, Regina ;
Eberle, Ute ;
Dangel, Alexandra ;
Treis, Bianca ;
Berger, Anja ;
Bengs, Katja ;
Fingerle, Volker ;
Liebl, Bernhard ;
Ackermann, Nikolaus ;
Sing, Andreas .
EUROSURVEILLANCE, 2020, 25 (09) :13-17
[8]   Early Transmission Dynamics in Wuhan, China, of Novel Coronavirus-Infected Pneumonia [J].
Li, Qun ;
Guan, Xuhua ;
Wu, Peng ;
Wang, Xiaoye ;
Zhou, Lei ;
Tong, Yeqing ;
Ren, Ruiqi ;
Leung, Kathy S. M. ;
Lau, Eric H. Y. ;
Wong, Jessica Y. ;
Xing, Xuesen ;
Xiang, Nijuan ;
Wu, Yang ;
Li, Chao ;
Chen, Qi ;
Li, Dan ;
Liu, Tian ;
Zhao, Jing ;
Liu, Man ;
Tu, Wenxiao ;
Chen, Chuding ;
Jin, Lianmei ;
Yang, Rui ;
Wang, Qi ;
Zhou, Suhua ;
Wang, Rui ;
Liu, Hui ;
Luo, Yinbo ;
Liu, Yuan ;
Shao, Ge ;
Li, Huan ;
Tao, Zhongfa ;
Yang, Yang ;
Deng, Zhiqiang ;
Liu, Boxi ;
Ma, Zhitao ;
Zhang, Yanping ;
Shi, Guoqing ;
Lam, Tommy T. Y. ;
Wu, Joseph T. ;
Gao, George F. ;
Cowling, Benjamin J. ;
Yang, Bo ;
Leung, Gabriel M. ;
Feng, Zijian .
NEW ENGLAND JOURNAL OF MEDICINE, 2020, 382 (13) :1199-1207
[9]   Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2 [J].
Lu, Renfei ;
Wu, Xiuming ;
Wan, Zhenzhou ;
Li, Yingxue ;
Zuo, Lulu ;
Qin, Jianru ;
Jin, Xia ;
Zhang, Chiyu .
VIROLOGICA SINICA, 2020, 35 (03) :344-347
[10]  
Mohon A. N., 2020, DEV VALIDATION DIREC, DOI [10.1101/2020.04.29.20075747, DOI 10.1101/2020.04.29.20075747]