Point-of-care (POC) testing techniques are essential in molecular diagnostics, particularly in resource-limited settings. The first step in molecular diagnostics is sample preparation which may include extraction of nucleic acids (NAs). Simple and efficient NA extraction is crucial for downstream applications such as pathogen detection, genetic analysis, disease screening and personalized medicine. However, traditional NA extraction methods are often labor-intensive, time consuming and require sophisticated equipment and training. Because they involve multiple steps, they are not easily scalable or adaptable for POC settings. Further, saliva and other non-invasive biofluids present unique challenges for nucleic acid extraction.
Researchers at the Biodesign Institute of Arizona State University had developed a novel nucleic acid extraction protocol (POC-Pure) designed for use at the point-of-care, leveraging a custom buffer-extraction utilizing silica to bind and release nucleic acids from salivary samples. This protocol eliminates multiple wash steps and is able to integrate into a low-cost, centrifuge-free microfluidic chip. It incorporates heat, proteinase K, and TCEP to overcome saliva's inherent challenges such as high RNase activity and viscosity, ensuring effective viral lysis and RNA preservation. Validated with SARS-CoV-2 and HPV samples, the method offers rapid, efficient extraction within 10 minutes and enables reliable amplification with minimal resource requirements.
This technique demonstrates robust and fast off- and on-chip NA extraction in biofluids with low detection limits to address many challenges faced with molecular POCT. Platforms like this could aid in disease detection and rapid responses at the point of care.
Potential Applications
- Point-of-care viral and bacterial salivary diagnostics (e.g., SARS-CoV-2, HPV)
- Rapid, low-cost molecular testing in resource-limited healthcare settings
- Integration into handheld or portable diagnostic devices
- Emergency and outbreak response testing platforms
- Home testing and remote patient monitoring solutions
Benefits and Advantages
- Significantly reduces extraction time by approximately 60%
- Eliminates multiple wash steps, simplifying the protocol
- Compatible with saliva samples by mitigating RNase activity and viscosity issues
- Cost-effective and centrifuge-free microfluidic chip integration
- Achieves clinically relevant low detection limits for viral nucleic acids
- Reduces waste volume by more than 50%
- Facilitates complete sample-to-answer diagnostic platforms at the point-of-care
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