Changes in the level of nucleic acids in blood may be correlated with some clinical disorders like cancer, stroke, trauma and autoimmune diseases, and thus, nucleic acids can serve as potential biomarkers for pathological processes. The requirement of technical equipment and operator expertise in effective information readout of modern molecular diagnostic technologies significantly restricted application outside clinical laboratories. The ability to detect nucleic acid biomarkers with off-the-shelf devices, which have the advantages of portability, simplicity, low cost and short response time, is critical to provide a prompt clinical result in circumstances where the laboratory instruments are not available. This review throws light on the current strategies and challenges for nucleic acid diagnosis with commercial portable devices, indicating the future prospect of portable diagnostic devices and making a great difference in improving the healthcare and disease surveillance in resource-limited areas.