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