CRISPR In Next-Generation Molecular Diagnostics

Nucleic acid testing, a key component of molecular diagnostics, is essential for understanding physiological processes and improving human and livestock health. This involves various techniques such as EIAs, PCR, RT-PCR, qPCR, FISH, and NGS, which, despite their high sensitivity and specificity, require sophisticated labs, costly equipment, significant time, and skilled personnel, limiting their use for quick point-of-care (POC) diagnostics. Emerging POC platforms like LAMP and RPA integrated with electrochemical optical biosensors address some of these limitations but often fall short in sensitivity. The CRISPR-Cas systems, especially the simpler Class 2 systems with single crRNA-binding proteins, offer a promising next-generation diagnostic approach. Technologies like NASBACC, DETECTER, and SHERLOCK utilize CRISPR-Cas systems to achieve highly sensitive, specific, and cost-effective diagnostics with single-nucleotide specificity, vital for detecting mutations linked to antibiotic or antiviral resistance.