Ultrafast image-based ELISA for sensitive detection of cytokines in allergen-induced asthmatic samples

Rapid measurement of cytokines, key mediators of immunity and inflammation is important for diagnosis of several inflammatory diseases including asthma, autoimmune and infectious diseases. Here, we report ultrafast detection of cytokines involving ELISA method by microwave irradiation in a domestic microwave oven in just 3 min 50 s. We have detected interleukin-5 (IL-5), interleukin-13 (IL-13) and interferon-ϒ (IFN-ϒ) by covalently immobilizing capture antibodies of IL-5, IL-13 and IFN-ϒ respectively, on activated polypropylene microtest plates (APPμTP) by microwave irradiation in 20 s followed by blocking, analyte (antigen) binding and detection molecule binding in 10 s, 100 s and 100 s respectively by microwave irradiation. The limit of detection (LOD) for IL-13 is 0.39 ng/ml, IL-5 is 0.078 ng/ml and IFN-ϒ is 0.125 ng/ml. The present method is further used for detection of IL-13 and IFN-ϒ in diseased asthmatic and control healthy mice samples. In case of IL-13, diseased samples show increased cytokine level compared to normal healthy samples; for IFN-ϒ, the result is reverse. As the present method for detection of cytokines is low cost, ultrafast, specific and sensitive, it can be used as an excellent substitute for conventional - ELISA based cytokine detection.