A team of researchers from the University of Florida has developed a new method that utilizes CRISPR technology for non-invasive blood tests to aid in the early diagnosis of cancer.
The approach demonstrates comparable effectiveness to the widely used reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assays and has the potential to be paired with a simple portable device for point-of-care clinical testing.
Detecting Cancer with MicroRNAs
The researchers focused on detecting microRNAs, small RNA molecules involved in regulating gene expression, which can serve as biomarkers for cancerous tumors.
However, previous clinical applications were limited due to the complexity of microRNAs and the lack of a sensitive detection tool.
The Role of Extracellular Vesicles
To overcome these challenges, the team targeted extracellular vesicles, nanosized particles released by cells that play crucial roles in cell functions and diseases.
Tumor cells release extracellular vesicles containing microRNAs associated with disease, making them a promising source for cancer markers.
Simplifying the Workflow with CRISPR
Taking advantage of CRISPR technology, the researchers aimed to develop a simplified workflow for detecting microRNAs.
Previous CRISPR tests involved multiple steps and were less sensitive than the gold standard, RT-qPCR. The team sought to streamline the process and minimize the risk of cross-contamination.
The “One Pot” Method
The team designed a “one pot” method, storing all necessary chemical agents in a single test tube, except for the sample.
Only the microRNA sample needs to be added to initiate the reaction. Surprisingly, this method showed comparable sensitivity to PCR tests while offering improved speed and efficiency.
Focus on Pancreatic Cancer
The researchers specifically targeted pancreatic cancer, a disease with high mortality rates and limited early detection methods. Early detection is challenging, often leading to late-stage diagnosis, reducing the chances of a cure.
The team aimed to develop biomarkers for pancreatic cancer diagnosis using the “EXTRA-CRISPR” method.
Game-Changing Potential
The “EXTRA-CRISPR” method represents a game-changing approach in the field of microRNA testing.
The team’s findings show promise for cancer diagnosis, particularly pancreatic cancer, when combined with robust microRNA biomarkers.
The method has the potential for point-of-care testing and can be coupled with a simple, low-cost portable device.
Advancing Point-of-Care Testing
As a proof of concept, the researchers adapted the method for two commonly used point-of-care detection methods.
They developed a portable smartphone-based device using readily available components, enabling fluorescence imaging of the test vials.
Additionally, they combined the EXTRA-CRISPR assay with a commercially available lateral-flow test strip to create an instrument-free point-of-care device.
Both methods yielded diagnostic results consistent with those obtained using a benchtop PCR machine.
Future Implications
The researchers have filed a patent application based on their work to facilitate the broad availability of the one-pot CRISPR assay and point-of-care technology for basic research and clinical use.
Further studies and developments are necessary to validate the safety and effectiveness of this method in a clinical setting, potentially revolutionizing cancer diagnosis and improving patient outcomes.
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The study was published in Nature Biomedical Engineering.
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