Advances in nucleic acid-based detection of dengue virus: From RT-PCR to CRISPR biosensing technologies

Dengue virus (DENV) is a global health concern, especially in tropical and subtropical regions. Rapid, cost-effective, and sensitive diagnostics are essential for point-of-care testing (POCT), genotyping, and surveillance. Current nucleic acid (NA)-based methods, such as RT-PCR, real-time RT-PCR, and isothermal techniques like RT-LAMP, are widely used but limited by cost, processing time, and equipment needs. Nanomaterial-based NA-biosensors have shown excellent potential for enhancing sensitivity, yet challenges remain in reproducibility, regulatory approval, and adaptability under variable field conditions. Recently, CRISPR-based diagnostics have gained attention for their high specificity, sensitivity, and rapid results, making them ideal for POCT. This review summarizes the evolution of NA-based DENV detection methods, from PCR and isothermal amplification to nanomaterial-based and CRISPR-based biosensors. CRISPR/Cas systems have achieved detection limits to picomolar, femtomolar, and attomolar levels, highlighting their potential for field-deployable diagnostics.