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Preprint in English | EuropePMC | ID: ppcovidwho-328671


An ultrasensitive assay for the detection of antibodies to SARS-CoV-2 is critically needed for evaluating the adaptive humoral immune response and infection rates in immunocompromised subpopulations. Here, we report an Ultrasensitive CRISPR-based Antibody Detection (UCAD) assay that translates the detection of serum antibodies against the receptor binding domain (RBD) of SARS-CoV-2 spike protein into CRISPR-based nucleic acid testing in a homogeneous solution and is thus 10,000 times more sensitive than the commercial immunoassay. The UCAD assay, which has been validated with 65 clinical anti-RBD-positive and 72 anti-RBD-negative sera collected from the general population, achieves 100% sensitivity and 97.2% specificity. We finally deployed UCAD to evaluate the levels of serum anti-RBD IgG and IgM in a cohort of 85 vaccinated kidney transplant recipients (KTRs), an especially vulnerable patient population with reported seroconversion rates of only 4-48%. Among the 85 vaccinated KTRs, UCAD successfully identified 68 seroconversion positive sera that were previously determined to contain “undetectable” levels of anti-SARS-CoV-2 using a clinical chemiluminescent immunoassay (CLIA) and has revealed significant differences in the levels of plasmablasts, type-2 T helper (Th2) cells, and type-17 T helper (Th17) cells between the UCAD-identified seroconversion positive and negative groups. As UCAD is a solution-based ultrasensitive assay that does not require specialized equipment or tedious operational and washing steps, we anticipate that it will find wide applications for clinical uses in both centralized laboratories and point-of-care settings.