Minimally instrumented SHERLOCK (miSHERLOCK) for CRISPR-based point-of-care diagnosis of SARS-CoV-2 and emerging variants

Helena de Puig; Rose A Lee; Devora Najjar; Xiao Tan; Luis R Soeknsen; Nicolaas M Angenent-Mari; Nina M Donghia; Nicole E Weckman; Audrey Ory; Carlos F Ng; Peter Q Nguyen; Angelo S Mao; Thomas C Ferrante; Geoffrey Lansberry; Hani Sallum; James Niemi; James J Collins

doi:10.1126/sciadv.abh2944

Minimally instrumented SHERLOCK (miSHERLOCK) for CRISPR-based point-of-care diagnosis of SARS-CoV-2 and emerging variants

Sci Adv. 2021 Aug 6;7(32):eabh2944. doi: 10.1126/sciadv.abh2944. Print 2021 Aug.

Authors

Helena de Puig^{1

2

3}, Rose A Lee^{1

4

5

6}, Devora Najjar^{1

2

7}, Xiao Tan^{1

2

3

6

8}, Luis R Soeknsen^{1

2

9}, Nicolaas M Angenent-Mari^{1

2}, Nina M Donghia¹, Nicole E Weckman¹, Audrey Ory^{1

10}, Carlos F Ng¹, Peter Q Nguyen¹, Angelo S Mao^{1

2}, Thomas C Ferrante¹, Geoffrey Lansberry¹, Hani Sallum¹, James Niemi¹, James J Collins^{11

2

3

9

10

12

13}

Affiliations

¹ Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA.
² Institute for Medical Engineering and Science, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
³ Infectious Disease and Microbiome Program, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
⁴ Division of Infectious Diseases, Department of Pediatrics, Boston Children's Hospital, Boston, MA 02115, USA.
⁵ Division of Infectious Diseases, Department of Medicine, Beth Israel Deaconess Medical Center, Boston, MA 02215, USA.
⁶ Harvard Medical School, Boston, MA 02115, USA.
⁷ MIT Media Lab, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
⁸ Division of Gastroenterology, Massachusetts General Hospital, Boston, MA 02114, USA.
⁹ Abdul Latif Jameel Clinic for Machine Learning in Health, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
¹⁰ College of Arts and Sciences, Harvard University, Cambridge, MA 02138, USA.
¹¹ Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA. [email protected].
¹² Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
¹³ Harvard-MIT Program in Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.

Abstract

The COVID-19 pandemic highlights the need for diagnostics that can be rapidly adapted and deployed in a variety of settings. Several SARS-CoV-2 variants have shown worrisome effects on vaccine and treatment efficacy, but no current point-of-care (POC) testing modality allows their specific identification. We have developed miSHERLOCK, a low-cost, CRISPR-based POC diagnostic platform that takes unprocessed patient saliva; extracts, purifies, and concentrates viral RNA; performs amplification and detection reactions; and provides fluorescent visual output with only three user actions and 1 hour from sample input to answer out. miSHERLOCK achieves highly sensitive multiplexed detection of SARS-CoV-2 and mutations associated with variants B.1.1.7, B.1.351, and P.1. Our modular system enables easy exchange of assays to address diverse user needs and can be rapidly reconfigured to detect different viruses and variants of concern. An adjunctive smartphone application enables output quantification, automated interpretation, and the possibility of remote, distributed result reporting.

Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

Abstract

Grants and funding