Pathogenic infection remains the primary threat to human health, such as the global COVID-19 pandemic. It is important to develop rapid, sensitive and multiplexed tools for detecting pathogens and their mutated variants, particularly the tailor-made strategies for point-of-care diagnosis allowing for use in resource-constrained settings. The rapidly evolving CRISPR/Cas systems have provided a powerful toolbox for pathogenic diagnostics via nucleic acid tests. In this review, we firstly describe the resultant promising class 2 (single, multidomain effector) and recently explored class 1 (multisubunit effector complexes) CRISPR tools. We present diverse engineering nucleic acid diagnostics based on CRISPR/Cas systems for pathogenic viruses, bacteria and fungi, and highlight the application for detecting viral variants and drug-resistant bacteria enabled by CRISPR-based mutation profiling. Finally, we discuss the challenges involved in on-site diagnostic assays and present emerging CRISPR systems and CRISPR cascade that potentially enable multiplexed and preamplification-free pathogenic diagnostics.
Keyphrases
- nucleic acid
- crispr cas
- genome editing
- drug resistant
- human health
- genome wide
- multidrug resistant
- single cell
- gram negative
- copy number
- regulatory t cells
- acinetobacter baumannii
- dendritic cells
- high throughput
- climate change
- antimicrobial resistance
- sars cov
- dna methylation
- pseudomonas aeruginosa
- type iii
- quantum dots
- sensitive detection
- genetic diversity