Field-deployable viral diagnostics using CRISPR-Cas13

被引：1087

作者：

Myhrvold, Cameron ^{[1
,2
]}

Freije, Catherine A. ^{[1
,2
,3
]}

Gootenberg, Jonathan S. ^{[1
,4
,5
,6
,7
]}

Abudayyeh, Omar O. ^{[1
,5
,6
,7
,8
]}

Metsky, Hayden C. ^{[1
,9
]}

Durbin, Ann F. ^{[3
,10
]}

Kellner, Max J. ^{[1
]}

Tan, Amanda L. ^{[11
]}

Paul, Lauren M. ^{[11
]}

Parham, Leda A. ^{[12
]}

Garcia, Kimberly F. ^{[12
]}

Barnes, Kayla G. ^{[1
,2
,13
]}

Chak, Bridget ^{[1
,2
]}

Mondini, Adriano ^{[14
]}

Nogueira, Mauricio L. ^{[15
]}

Isern, Sharon ^{[11
]}

Michael, Scott F. ^{[11
]}

Lorenzana, Ivette ^{[12
]}

Yozwiak, Nathan L. ^{[1
,2
]}

MacInnis, Bronwyn L. ^{[1
,13
]}

Bosch, Irene ^{[10
,16
]}

Gehrke, Lee ^{[3
,10
,17
]}

Zhang, Feng ^{[1
,2
,5
,6
,7
]}

Sabeti, Pardis C. ^{[1
,2
,3
,13
,18
]}

机构：

[1] Broad Inst Massachusetts Inst Technol MIT & Harva, Cambridge, MA 02142 USA

[2] Harvard Univ, Dept Organismal & Evolutionary Biol, Ctr Syst Biol, Cambridge, MA 02138 USA

[3] Harvard Med Sch, Div Med Sci, PhD Program Virol, Boston, MA 02115 USA

[4] Harvard Med Sch, Dept Syst Biol, Boston, MA 02115 USA

[5] MIT, McGovern Inst Brain Res, Cambridge, MA 02139 USA

[6] MIT, Dept Brain & Cognit Sci, Cambridge, MA 02139 USA

[7] MIT, Dept Biol Engn, Cambridge, MA 02139 USA

[8] MIT, Dept Hlth Sci & Technol, Cambridge, MA 02139 USA

[9] MIT, Dept Elect Engn & Comp Sci, Cambridge, MA 02139 USA

[10] MIT, Inst Med Engn & Sci, Cambridge, MA 02139 USA

[11] Florida Gulf Coast Univ, Dept Biol Sci, Ft Myers, FL 33965 USA

[12] Univ Nacl Autonoma Honduras, Inst Invest Microbiol, Ctr Invest Genet, Tegucigalpa, Honduras

[13] Harvard Sch Publ Hlth, Dept Immunol & Infect Dis, Boston, MA 02115 USA

[14] Sao Paulo State Univ, Sch Pharmaceut Sci, Araraquara Lab Publ Hlth, Sao Paulo, Brazil

[15] Fac Med Sao Jose do Rio Preto, Lab Pesquisas Virol, Sao Paulo, Brazil

[16] Mt Sinai Sch Med, Dept Med, New York, NY 10029 USA

[17] Harvard Med Sch, Dept Microbiol & Immunobiol, Boston, MA 02115 USA

[18] HHMI, Chevy Chase, MD 20815 USA

来源：

SCIENCE | 2018年 / 360卷 / 6387期

基金：

巴西圣保罗研究基金会;

关键词：

ZIKA-VIRUS; PCR; AMPLIFICATION; INFECTION; AMERICA; ASSAY;

D O I：

10.1126/science.aas8836

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Mitigating global infectious disease requires diagnostic tools that are sensitive, specific, and rapidly field deployable. In this study, we demonstrate that the Cas13-based SHERLOCK (specific high-sensitivity enzymatic reporter unlocking) platform can detect Zika virus (ZIKV) and dengue virus (DENV) in patient samples at concentrations as low as 1 copy per microliter. We developed HUDSON (heating unextracted diagnostic samples to obliterate nucleases), a protocol that pairs with SHERLOCK for viral detection directly from bodily fluids, enabling instrument-free DENVdetection directly from patient samples in <2 hours. We further demonstrate that SHERLOCK can distinguish the four DENV serotypes, as well as region-specific strains of ZIKV from the 2015-2016 pandemic. Finally, we report the rapid (<1 week) design and testing of instrument-free assays to detect clinically relevant viral single-nucleotide polymorphisms.

引用

页码：444 / 448

页数：5

共 26 条

[1] C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector [J].

Abudayyeh, Omar O. ;

Gootenberg, Jonathan S. ;

Konermann, Silvana ;

Joung, Julia ;

Slaymaker, Ian M. ;

Cox, David B. T. ;

Shmakov, Sergey ;

Makarova, Kira S. ;

Semenova, Ekaterina ;

Minakhin, Leonid ;

Severinov, Konstantin ;

Regev, Aviv ;

Lander, Eric S. ;

Koonin, Eugene V. ;

Zhang, Feng .

SCIENCE, 2016, 353 (6299)

[2] Value of Routine Dengue Diagnostic Tests in Urine and Saliva Specimens [J].

Andries, Anne-Claire ;

Duong, Veasna ;

Ly, Sowath ;

Cappelle, Julien ;

Kim, Kim Srorn ;

Try, Patrich Lorn ;

Ros, Sopheaktra ;

Ong, Sivuth ;

Huy, Rekol ;

Horwood, Paul ;

Flamand, Marie ;

Sakuntabhai, Anavaj ;

Tarantola, Arnaud ;

Buchy, Philippe .

PLOS NEGLECTED TROPICAL DISEASES, 2015, 9 (09)

[3] Antibody-based assay discriminates Zika virus infection from other flaviviruses [J].

Balmaseda, Angel ;

Stettler, Karin ;

Medialdea-Carrera, Raquel ;

Collado, Damaris ;

Jin, Xia ;

Zambrana, Jose Victor ;

Jaconi, Stefano ;

Cameroni, Elisabetta ;

Saborio, Saira ;

Rovida, Francesca ;

Percivalle, Elena ;

Ijaz, Samreen ;

Dicks, Steve ;

Ushiro-Lumb, Ines ;

Barzon, Luisa ;

Siqueira, Patricia ;

Brown, David W. G. ;

Baldanti, Fausto ;

Tedder, Richard ;

Zambon, Maria ;

de Filippis, A. M. Bispo ;

Harris, Eva ;

Corti, Davide .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2017, 114 (31) :8384-8389

[4] Rapid antigen tests for dengue virus serotypes and Zika virus in patient serum [J].

Bosch, Irene ;

de Puig, Helena ;

Hiley, Megan ;

Carre-Camps, Marc ;

Perdomo-Celis, Federico ;

Narvaez, Carlos F. ;

Salgado, Doris M. ;

Senthoor, Dewahar ;

O'Grady, Madeline ;

Phillips, Elizabeth ;

Durbin, Ann ;

Fandos, Diana ;

Miyazaki, Hikaru ;

Yen, Chun-Wan ;

Gelvez-Ramirez, Margarita ;

Warke, Rajas V. ;

Ribeiro, Lucas S. ;

Teixeira, Mauro M. ;

Almeida, Roque P. ;

Munoz-Medina, Jose E. ;

Ludert, Juan E. ;

Nogueira, Mauricio L. ;

Colombo, Tatiana E. ;

Terzian, Ana C. B. ;

Bozza, Patricia T. ;

Calheiros, Andrea S. ;

Vieira, Yasmine R. ;

Barbosa-Lima, Giselle ;

Vizzoni, Alexandre ;

Cerbino-Neto, Jose ;

Bozza, Fernando A. ;

Souza, Thiago M. L. ;

Trugilho, Monique R. O. ;

de Filippis, Ana M. B. ;

de Sequeira, Patricia C. ;

Marques, Ernesto T. A. ;

Magalhaes, Tereza ;

Diaz, Francisco J. ;

Restrepo, Berta N. ;

Marin, Katerine ;

Mattar, Salim ;

Olson, Daniel ;

Asturias, Edwin J. ;

Lucera, Mark ;

Singla, Mohit ;

Medigeshi, Guruprasad R. ;

de Bosch, Norma ;

Tam, Justina ;

Gomez-Marquez, Jose ;

Clavet, Charles .

SCIENCE TRANSLATIONAL MEDICINE, 2017, 9 (409)

[5] Rapid and specific detection of Asian- and African-lineage Zika viruses [J].

Chotiwan, Nunya ;

Brewster, Connie D. ;

Magalhaes, Tereza ;

Weger-Lucarelli, James ;

Duggal, Nisha K. ;

Ruckert, Claudia ;

Chilinh Nguyen ;

Luna, Selene M. Garcia ;

Fauver, Joseph R. ;

Andre, Barb ;

Gray, Meg ;

Black, William C. ;

Kading, Rebekah C. ;

Ebel, Gregory D. ;

Kuan, Guillermina ;

Balmaseda, Angel ;

Jaenisch, Thomas ;

Marques, Ernesto T. A. ;

Brault, Aaron C. ;

Harris, Eva ;

Foy, Brian D. ;

Quackenbush, Sandra L. ;

Perera, Rushika ;

Rovnak, Joel .

SCIENCE TRANSLATIONAL MEDICINE, 2017, 9 (388)

[6] Coupling Sensitive Nucleic Acid Amplification with Commercial Pregnancy Test Strips [J].

Du, Yan ;

Pothukuchy, Arti ;

Gollihar, Jimmy D. ;

Nourani, Armin ;

Li, Bingling ;

Ellington, Andrew D. .

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2017, 56 (04) :992-996

[7] Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection [J].

East-Seletsky, Alexandra ;

O'Connell, Mitchell R. ;

Knight, Spencer C. ;

Burstein, David ;

Cate, Jamie H. D. ;

Tjian, Robert ;

Doudna, Jennifer A. .

NATURE, 2016, 538 (7624) :270-+

[8] Rapid molecular diagnostic test for Zika virus with low demands on sample preparation and instrumentation [J].

Eboigbodin, Kevin E. ;

Brummer, Mirko ;

Ojalehto, Tuomas ;

Hoser, Mark .

DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE, 2016, 86 (04) :369-371

[9] Establishment and cryptic transmission of Zika virus in Brazil and the Americas [J].

Faria, N. R. ;

Quick, J. ;

Claro, I. M. ;

Theze, J. ;

de Jesus, J. G. ;

Giovanetti, M. ;

Kraemer, M. U. G. ;

Hill, S. C. ;

Black, A. ;

da Costa, A. C. ;

Franco, L. C. ;

Silva, S. P. ;

Wu, C. -H. ;

Raghwani, J. ;

Cauchemez, S. ;

du Plessis, L. ;

Verotti, M. P. ;

de Oliveira, W. K. ;

Carmo, E. H. ;

Coelho, G. E. ;

Santelli, A. C. F. S. ;

Vinhal, L. C. ;

Henriques, C. M. ;

Simpson, J. T. ;

Loose, M. ;

Andersen, K. G. ;

Grubaugh, N. D. ;

Somasekar, S. ;

Chiu, C. Y. ;

Munoz-Medina, J. E. ;

Gonzalez-Bonilla, C. R. ;

Arias, C. F. ;

Lewis-Ximenez, L. L. ;

Baylis, S. A. ;

Chieppe, A. O. ;

Aguiar, S. F. ;

Fernandes, C. A. ;

Lemos, P. S. ;

Nascimento, B. L. S. ;

Monteiro, H. A. O. ;

Siqueira, I. C. ;

de Queiroz, M. G. ;

de Souza, T. R. ;

Bezerra, J. F. ;

Lemos, M. R. ;

Pereira, G. F. ;

Loudal, D. ;

Moura, L. C. ;

Dhalia, R. ;

Franca, R. F. .

NATURE, 2017, 546 (7658) :406-+

[10] One-step RT-PCR for detection of Zika virus [J].

Faye, Oumar ;

Faye, Ousmane ;

Dupressoir, Anne ;

Weidmann, Manfred ;

Ndiaye, Mady ;

Sall, Amadou Alpha .

JOURNAL OF CLINICAL VIROLOGY, 2008, 43 (01) :96-101

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