In a groundbreaking new study, researchers have improved the effectiveness and efficiency of CRISPR diagnostic technology, paving the way for faster, more sensitive and deployable molecular diagnosis. The investigation, published in Nature Communications, reveals new potential for simple and sensitive nucleic acid detection in various diagnostic settings using a new and improved clustered regularly interspaced short palindromic repeats (CRISPR) technology.
PCR-based nucleic acid detection methods have long been considered the gold standard due to their high sensitivity and specificity. However, there is a growing need for alternative solutions that are fast, cost-effective and easy to use. CRISPR technology is emerging as a powerful tool for nucleic acid detection due to its affordability and simplicity.
The existing CRISPR method requires two steps: a preamplification step of the target nucleic acids and a detection of the CRISPR enzymes. While this method is simpler and more effective than standard PCR-based methods, the two-step requirement limits practical application and lacks quantitative detection capability.
Changchun Liu, professor of Biomedical Engineering at UConn Health, has developed an asymmetric transcleavage behavior of competitive crRNAs in the CRISPR-Cas12a reaction. Based on this finding, Liu’s team developed a sensitive, amplification-free asymmetric CRISPR assay to quantitatively detect nucleic acids.
Using CRISPR-Cas12a, an RNA-guided DNA enzyme that can be harnessed for gene editing, the researchers developed a new CRISPR assay by taking advantage of a unique competitive reaction between a full-length crRNA and a cleaved crRNA. The reaction induces signal amplification significantly improving target detection signal sensitivity.
This improved diagnostic technology achieved atomolar detection sensitivity—1,000 times more sensitive than conventional CRISPR detection—without requiring preamplification. To test its effectiveness in clinical settings, the researchers applied it to analyze biomarker microRNA-19a in plasma samples from patients with bladder cancer successfully demonstrating its potential as a powerful tool for simple liquid biopsy—a fast and sensitive cancer screening technique.
Jeong Moon, postdoctoral researcher in Liu’s laboratory says “We envision that it could have broad clinical implications in early cancer diagnosis.”
With these advancements in CRISPR diagnostic technology showing great promise across various applications including early cancer diagnosis and infectious disease detection; research continues to push boundaries towards better healthcare outcomes.
According to source.
This article was originally published on Phys.org