Source – Forbes
Introduction to the Study
At the 102nd General Session of the International Association for Dental Research (IADR), a groundbreaking study on rapid oral pathogen detection using CRISPR-based diagnostics was presented. The study, titled “Rapid Specific Detection of Oral Pathogens Using CRISPR-Based Diagnostics,” aimed to develop a cost-effective and swift technique for the widespread detection and screening of oral microorganisms, suitable for point-of-care settings.
Innovative Approach and Findings
Led by Batbileg Bor of the ADA Forsyth Institute, the study tailored the CRISPR-Cas-based diagnostic platform Specific High-Sensitivity Enzymatic Reporter Unlocking (SHERLOCK) for species-specific detection of oral bacterial pathogens and human papillomavirus (HPV) nucleic acids. By developing a computational pipeline to generate guide-RNAs and species-specific gene primers, the investigators synthesized constructs using cell-free biosynthesis systems. Experimental validation demonstrated the constructs’ specificity and sensitivity through fluorescence readings of reporter RNAs.
The study successfully detected oral bacteria at the single-molecule level, maintaining specificity even in the presence of off-target DNA in saliva. Moreover, the assay was refined to detect common oral pathogens directly from unprocessed saliva samples, with results aligning fully with other detection methods when tested on patient saliva samples. Additionally, the investigators specifically detected various HPV strains from genomic DNA backgrounds, showcasing the versatility of the method.
Implications and Future Directions
The developed method holds immense promise for scalable implementation at point-of-care settings, as it offers rapid detection within approximately 35 minutes, minimal cost, and requires no specialized skills or techniques. Given SHERLOCK’s specificity for nucleic acid sequences, future assays can be devised to detect other microorganisms, such as fungi and archaea, as well as human gene products.
In summary, the study’s innovative approach to oral pathogen detection using CRISPR-based diagnostics represents a significant advancement in dental research. By offering a rapid, cost-effective, and highly specific detection method, this technique has the potential to revolutionize oral healthcare and facilitate early intervention strategies for oral diseases.
Also Read: RNA Interference vs. CRISPR-Cas9: A Battle for Gene Silencing Supremacy
