This protocol describes a novel ultrasound-mediated ELISA procedure that can dramatically reduce the ELISA timing to 40 minutes without losing its specificity or sensitivity. Ultrasound-mediated ELISA was best achieved on an activated microtiter plate which was able to covalently bind antigen or antibody. Binding occured in 10 min when subjected to ultrasound waves in a sonicator bath, operating at an output power of 120 W. Blocking, antibody binding and secondary antibody–enzyme conjugate binding were also accomplished in 10 min each in the sonicator bath under similar conditions. Total IgE detection by 40 min-SELISA method gives similar absorbance value to that obtained by 20 h- conventional procedure. In fact any type of ELISA can be carried out by SELISA technique. SELISA method is sensitive and can detect lower concentration of analyte that could be significantly useful to confirm false negative cases. In short, SELISA procedure is rapid, sensitive, specific and reproducible; hence better option than conventional ELISA procedure.