pISSN 1225-7117 eISSN 2288-8268

CRISPR/Cas-based diagnostics before COVID-19 outbreak

Cas type System name1 Target Amplification2 Time LOD Sensitivity / Specificity Readout3 Instrument Ref.
Cas9 NASBACC ZIKA virus Isothermal RNA amplification (NASBA) ~3 h 1.8 cps/μL fM/1 nt4 C(chlorophenol red-β-D-galactopyranoside) Portable electronic optical reader [53]
dCas9+PCR Mycobacterium tuberculosis PCR 10 min after PCR - One copy/- B(Luciferase) - [58]
CRISPREXPAR Listeria monocytogenes EXPAR < 1 h 0.82 amol aM/1 nt F(SYBR Green 1) - [60]
RCH (dCas9-split HRB, RCA) miRNAs (NSCLC) RCA < 4 h - fM/1 nt C(TMB) - [42]
Cas13a SHELOCK ZIKA virus RPA 2 - 5 h - aM/1 nt F(FAM) - [5]
SHELOCKv2 Genomic DNA RPA 0.5 - 3 h - zM/1 nt F(FAM, TEX, Cy5, HEX); C (Gold-NP, anti-FAM Abs) ) - [54]
HUDSON+SHELOCK ZIKA virus RPA < 2 h - aM/1 nt F(FAM); C(Gold-NP, anti-FAM Abs) - [57]
Cas12a HOLMES Genomic DNA PCR; RT-PCR < 1 h - aM/1 nt F(HEX) - [61]
DETECTR HPV RPA ~2 h - aM/6 nt F(FAM) - [7]
Cas12b HOLMESv2 Genomic DNA LAMP; RTLAMP; Asymmetric PCR ~1 h - aM/1 nt F(HEX, FAM) - [55]

1NASBACC, Nucleic Acid Sequence-Based Amplification-CRISPR Cleavage; CRISRP-EXPAR, CRISPR-cas9 triggered isothermal EXponential Amplification Reaction; RCH, Rolling Circle Amplification-CRISPR-split-HRP; SHELOCK, Specific High-Sensitivity Enzymatic Reporter UnLOCKing; HUDSON, Heating Unextracted Diagnostic Sampled to Obliterate Nuclease; HOLMES, one-HOur low-cost Multipurpose highly Efficient System; DETECTR, DNA endonuclease-targeted CRISPR trans reporter, 2EXPAR, EXponential Amplification Reaction; RCA, Rolling Circle Amplification; RPA, Recombinant Polymerase Amplification; RT-RPA; Reverse transcriptional-RPA; LAMP, Loop-Mediated Isothermal Amplification; RT-LAMP, Reverse transcriptional-LAMP, 3C, colorimetric; B, bioluminescent; F, fluorescent, 4nt, nucleotide

Korean Society for Biotechnology and Bioengineering Journal 2023;38:77~89 https://doi.org/10.7841/ksbbj.2023.38.2.77
© Korean Society for Biotechnology and Bioengineering