Key result
The tandem Cas13a/crRNA-mediated CRISPR-FET biosensor achieved a detection limit of 1.56 aM for direct viral gene sensing and successfully distinguished HCV-infected patients from normal controls.
Why the study?
Ultrasensitive and timely pathogen identification without PCR has remained a long-lasting challenge in translating field-effect transistor applications from the laboratory to the clinic.
The CRISPR-FET biosensor provides an ultrasensitive, amplification-free method for detecting viral RNA with potential for scalable molecular diagnostics.
May support preclinical viral diagnostics; leaves open clinical translation pending human validation.
The path toward field-effect transistor (FET) application from laboratory to clinic has delivered a compelling push in the biomedical domain, yet ultrasensitive and timely pathogen identification without PCR remains a long-lasting challenge. Herein, we create a generic check station termed "CRISPR-FET", first incorporating the CRISPR/Cas13a system within the FET modality, for accelerated and unamplified detection of viral RNA. Unlike conventional FETs bearing target-specific receptors, this sensor holds three unique advancements: (i) an ingenious sensing mechanism is used, which converts the signal of a large-sized analyte into an on-chip cleavage response of an immobilized CRISPR reporter, enabling signal generation events to occur all within the Debye length; (ii) the multipurpose inspection of the CoV ORF1ab, CoV N gene, and HCV RNA unveils the potential for "one-for-all" scalable FET-based molecular diagnostics; and (iii) it is shown that Cas13a-crRNAs targeting different sites of the viral genome can be deployed in tandem to amplify the FET response, empowering the detection limit down to 1.56 aM, which is a world-record level of sensitivity in the FET for direct viral gene sensing. Notably, a brilliant clinical applicability was made in distinguishing HCV-infected patients from normal controls. Overall, this study sheds new insights into FET-based nucleic acid sensing technology and invokes a vision for its possible future roles in diagnosis of various viral diseases.
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Li et al. (2022) studied Viral infection (HCV, CoV). CRISPR-FET biosensor vs. Normal controls was evaluated on Detection limit of viral RNA. The tandem Cas13a/crRNA-mediated CRISPR-FET biosensor achieved a detection limit of 1.56 aM for direct viral gene sensing and successfully distinguished HCV-infected patients from normal controls.
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