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Abstract
CRISPR diagnostics are powerful tools for detecting nucleic acids but are generally not deployable for the detection of clinically important proteins. Here, we report an ultrasensitive CRISPR-based antibody detection (UCAD) assay that translates the detection of anti-SARS-CoV-2 antibodies into CRISPR-based nucleic acid detection in a homogeneous solution and is 10,000 times more sensitive than the classic immunoassays. Clinical validation using serum samples collected from the general population (n = 197), demonstrates that UCAD has 100% sensitivity and 98.5% specificity. With ultrahigh sensitivity, UCAD enables the quantitative analysis of serum anti-SARS-CoV-2 levels in vaccinated kidney transplant recipients who are shown to produce “undetectable” anti-SARS-CoV-2 using standard immunoassay. Because of the high sensitivity and simplicity, we anticipate that, upon further clinical validation against large cohorts of clinical samples, UCAD will find wide applications for clinical uses in both centralized laboratories and point-of-care settings.
CRISPR diagnostics are routinely used for the detecting nucleic acids, but rarely for clinically important proteins. Here, by translating a CRISPR-based DNA test into an ultrasensitive assay for antibodies, the authors achieve antibody detection from serum samples at attomolar concentrations.
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1 Sichuan University, Key Laboratory of Green Chemistry & Technology of Ministry of Education, College of Chemistry, Analytical & Testing Center, Chengdu, China (GRID:grid.13291.38) (ISNI:0000 0001 0807 1581)
2 Sichuan University, Urology Department, Urology Research Institute, Organ Transplantation Center, West China Hospital, Chengdu, China (GRID:grid.13291.38) (ISNI:0000 0001 0807 1581)
3 Sichuan University, Department of Laboratory Medicine, West China Hospital, Chengdu, China (GRID:grid.13291.38) (ISNI:0000 0001 0807 1581)
4 Second Hospital Affiliated to Southern University of Science and Technology, Shenzhen Key Laboratory of Pathogen and Immunity, Shenzhen Third People’s Hospital, Shenzhen, China (GRID:grid.263817.9) (ISNI:0000 0004 1773 1790)
5 the First Affiliated Hospital of Guangzhou Medical University, State Key Laboratory of Respiratory Disease, National Clinical Research Centre for Respiratory Disease, Guangzhou Institute of Respiratory Health, Guangzhou, China (GRID:grid.470124.4)
6 Sichuan University, Key Laboratory of Green Chemistry & Technology of Ministry of Education, College of Chemistry, Analytical & Testing Center, Chengdu, China (GRID:grid.13291.38) (ISNI:0000 0001 0807 1581); Brock University, St. Catharines, Department of Chemistry, Centre for Biotechnology, Ontario, Canada (GRID:grid.411793.9) (ISNI:0000 0004 1936 9318)