Field-deployable viral diagnostics using CRISPR-Cas13

被引：1077

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

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