Creating an improved workflow for paper-based malaria diagnostics by integrating total lysis of whole blood

Detection of parasite biomarkers is critical for diagnosis and treatment monitoring; however, proper analysis of whole blood samples typically requires red blood cell lysis. While lysis is easily achieved in laboratory environments, there is a clinical need for low-cost technology to collect, process, and analyze these samples in low-resource settings. Dried blood spot cards offer a simple way to microsample blood in these settings, but standard cards cannot perform sample preprocessing, including the required lysis step. To meet this need, we have developed a three-dimensional paper-based lysis device that was fabricated alternatively by laser cutting and by wax-printing. This device effectively achieved 100% cellular lysis independent of sample composition. Successful lysis from these devices resulted in the detection of a malaria biomarker comparable to the standard liquid lysis procedure of Plasmodium-infected samples. Moreover, if device-dried, samples displayed antigen stability for over 1 week at room temperature, which facilitated delayed laboratory testing as well. The results show that these lysis devices have the potential to expand the types of environments from which samples tested for malaria can be collected.