Field-deployable viral diagnostics using CRISPR-Cas13

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Myhrvold, Cameron ^{[1, 2]} ; Freije, Catherine A. ^{[1, 2, 3]} ; Gootenberg, Jonathan S. ^{[4, 1, 5, 6, 7]} ; Abudayyeh, Omar O. ^{[4, 1, 6, 7, 8]} ; Metsky, Hayden C. ^{[9, 1]} ; Durbin, Ann F. ^{[10, 3]} ; 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 ^{[10, 17, 3]} ; Zhang, Feng ^{[4, 1, 6, 7, 2]} ; Sabeti, Pardis C. ^{[1, 2, 3, 13, 18]}

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^[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] MIT, Dept Biol Engn, Cambridge, MA 02139 - USA

^[5] Harvard Med Sch, Dept Syst Biol, Boston, MA 02115 - USA

^[6] MIT, McGovern Inst Brain Res, Cambridge, MA 02139 - USA

^[7] MIT, Dept Brain & Cognit Sci, 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

• Tipo de documento: Artigo Científico
• Fonte: Science; v. 360, n. 6387, p. 444-448, APR 27 2018.
• Citações Web of Science: 82

Resumo

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. (AU)

• Processo FAPESP: 13/21719-3 - Estudo epidemiológico da dengue (sorotipos 1 a 4) em coorte prospectiva de São José do Rio Preto, São Paulo, Brasil, durante 2014 a 2018
• Beneficiário: Maurício Lacerda Nogueira
• Modalidade de apoio: Auxílio à Pesquisa - Temático