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| Image created by Dr. Michael J. Miller |
A team of researchers from the Hefei Institutes of Physical Science of the Chinese Academy of Sciences has developed a point-of-care testing (POCT) system based on microfluidic CRISPR-Dx technology, significantly improving the sensitivity, speed, and multiplexing capability of pathogen detection.
The findings were published in Biosensors and Bioelectronics.
CRISPR/Cas-based molecular diagnostics can precisely detect specific gene sequences in pathogens, including genetic mutations and abnormalities, providing important information for disease diagnosis and treatment.
Although it offers high accuracy and is considered a promising alternative to polymerase chain reaction, it has limitations like low sensitivity, limited capacity, and the need for multiple lab instruments, making it challenging for rapid and convenient multi-pathogen detection in point-of-care settings.
To address these challenges, the researchers combined microfluidic chips with CRISPR/Cas12a technology to create a dual-stage system that separates the amplification and detection processes. This separation, along with a two-stage centrifugation method, prevents contamination during testing.
The system also allows for customizable detection channels based on the number of pathogens, improving sensitivity and the ability to test multiple pathogens at once.
The Lift-CM POCT analysis system, which includes a heating platform, centrifugation module, and real-time fluorescence detection. It is compact, easy to use, and controlled by a smartphone APP, reducing the need for multiple lab instruments. This makes CRISPR-Dx technology more practical in resource-limited settings.
"Lift-CM is faster and easier to use than traditional CRISPR methods," said Dr. Zhu Cancan, a team member. "It's great for accurately detecting viruses like influenza and Ebola."
The entire process, from amplification to detection, takes just 30 minutes, making it a fast and effective solution for pathogen detection in emergency situations, disease screening, and home health monitoring.
This breakthrough offers new hope for early diagnosis, faster response times, and ultimately, better health care outcomes worldwide.
Reference
Taowei Shu et al, Lift-CM: An integrated lift-heater centrifugal microfluidic platform for point-of-care pathogen nucleic acid detection using isothermal amplification and CRISPR/Cas12a, Biosensors and Bioelectronics (2025). DOI: 10.1016/j.bios.2025.117178
Abstract
Pathogen nucleic acid detection technology based on isothermal amplification and CRISPR/Cas12a system offers advantages in terms of high sensitivity, high specificity, and rapidity. However, this method has not been widely applied because of its shortcomings in utilizing conventional instruments, which cannot satisfy the requirements for Point of Care Testing (POCT), such as integration, convenience, and miniaturization. In this study, we developed an integrated lift-heater centrifugal microfluidic platform (Lift-CM) to automate the processes of isothermal amplification and CRISPR/Cas12a detection. A spatially encoded centrifugal microfluidic disc (SEC-disc) was employed to physically separate the amplification and detection processes while expanding the number of targets. The design of the dual-temperature and lift-heating centrifugal mechanism of the Lift-CM platform ensures that there is no manual intervention during amplification and detection processes. A smartphone-based app enables the setting of key parameters and monitoring of the experimental process, presenting results through a generated report that includes real-time fluorescence curves. We analyzed the plasmids of the Crimean-Congo hemorrhagic fever virus, Ebola virus, and five influenza viruses using different amplification methods (RPA/LAMP) to demonstrate the good compatibility of the Lift-CM platform with various amplification schemes. In clinical validation, the detection of H3N2-positive samples was completed within 30 min, and the results were highly consistent with qPCR results. This portable and compact platform offers a novel alternative solution for both clinical and at-home pathogen nucleic acid detection in the future.