Rapid Elisa from Whole Blood in A Microcentrifuge Tube

Herein, we describe an enzyme-linked immunosorbent assay (ELISA) technique carried out in a polypropylene microcentrifuge tube preactivated by a photolinker, 1-fluoro-2-nitro-4-azidobenzene (FNAB) by a photochemical reaction. Immobilization of capture molecule (antibody) and BSA-blocking are carried out in the tube by ultrasound waves in a sonicator bath in 10 min for each step. Around one drop (100 μl) of blood is directly collected into the tube. By the time, the sample reaches laboratory (nearly 1 h), the analyte present in blood binds to the capture molecule already present in the tube. That means only one step (antibody-conjugate binding step) has to be carried out requiring only 10 min by ultrasound waves. The advantage of the method is that sample collection and testing can be done in the same tube and more importantly without the need of getting serum by a time consuming process. Also, time is saved for Ag-Ab binding step making the procedure more rapid.