To date, 55% of inhabitants of the developing world still live in rural regions and they have a very urgent need to improve the level of disease diagnosis, especially in resource-limited settings. Herein, we present a microfluidic system that centrifuges whole blood and quantifies contained biomarkers in a rapid, feasible and integrated way. Such a device provides a blood-in/answer-out capability and is small enough to be carried by any individual in any environment. The successful integration a hand-powered centrifuge and immunoassay unit within a rotational paper-based device allows for diagnostic application by untrained users and in environments where access to electricity cannot be assumed. In addition, the low cost (~$ 0.5), light weight and small instrumental footprint make the device ideally suited for rapid on-site detection. To validate the applicability of the system in a clinical diagnostic testing, we successfully perform enzyme-linked immunosorbent assay (ELISA) analysis of carcinoembryonic antigen and alpha fetoprotein from human blood samples. We expect that this powerful platform technology will provide the opportunities for point-of-care diagnosis in resource-limited settings.