The groundbreaking research led by bioengineering professor and Dean of The Grainger College of Engineering, Rashid Bashir, has resulted in the development of new technology that provides rapid and highly sensitive detection of multi-drug-resistant bacteria and other pathogens at low concentrations. This innovative technology was recently featured in an article in the prestigious Proceedings of the National Academy of Sciences (PNAS).
The research team designed a CRISPR-based test that can swiftly detect minimal levels of pathogen genetic material in blood without the need for nucleic acid amplification. In traditional CRISPR/Cas-based diagnostic tests, guide RNAs bind to pathogen DNA or RNA, activating Cas enzymes to cleave reporter nucleic acids that fluoresce upon cleavage. However, these tests often struggle to detect pathogens at low levels without a preamplification step.
Bashir’s team overcame this limitation by creating a CRISPR-based diagnostic test that eliminates the need for amplification. They achieved this by combining two CRISPR/Cas units into a complex called CRISPR-Cascade. One unit contains a guide RNA specific to a chosen pathogen nucleic acid and a Cas protein. When the Cas cleaves engineered nucleic acids added to the system, certain parts of the nucleic acids can bind and activate a second CRISPR/Cas unit, initiating a positive feedback loop with a high signal-to-noise ratio.
The test demonstrated exceptional sensitivity, detecting multi-drug-resistant Staphylococcus aureus DNA at concentrations significantly lower than those detected by a single Cas test. Additionally, the test accurately provided a simple “yes/no” result for the presence of any one pathogen in samples spiked with four common bloodstream pathogens.
The implications of this research are significant, as it paves the way for the development of highly sensitive CRISPR-based diagnostic tests capable of detecting pathogens within minutes without the need for nucleic acid amplification. This technology has the potential to revolutionize the field of pathogen detection and significantly impact the diagnosis and treatment of infectious diseases.
For more information on this groundbreaking research, you can refer to the article published in the Proceedings of the National Academy of Sciences titled “Amplification-free, OR-gated CRISPR-Cascade reaction for pathogen detection in blood samples.” The study was conducted by Jongwon Lim et al. and can be accessed with the DOI 10.1073/pnas.2420166122.
This research was made possible by the University of Illinois Grainger College of Engineering, showcasing their commitment to innovation and advancing scientific knowledge. For more information on their work, visit their official website.
In conclusion, the development of this new CRISPR-based diagnostic test represents a significant advancement in the field of pathogen detection, offering a faster and more sensitive method for identifying harmful bacteria and viruses in biological samples. This technology has the potential to revolutionize diagnostic practices and improve patient outcomes in the fight against infectious diseases.
