Peanut allergy is life threatening health concern that caused by allergenic proteins termed as Arachis hypogea that present in peanut. The most fatal allergic reaction to peanuts is anaphylaxis that is harmful to the individuals who are allergic to peanut allergens. Sometimes, traces of peanut allergens contamination are also occurred during food processing. Hence, it is very much important to detect peanut allergen in food products and our proposed study was carried out to detect peanut allergens (Arachis hypogea allergen) in various processed and unprocessed foods using real-time Polymerase Chain reaction (real-time PCR) technique as a rapid and sensitive method. First, DNA was extracted from the food samples using commercially available DNA extraction kit and further quantity & quality of the extracted DNA were tested. The extracted DNA was further observed for amplification to test the presence of peanut allergens in chosen food samples to detect Arachis hypogea allergen (peanut allergen) positive food samples. The sensitivity and specificity of proposed method was assayed for its limit of detection which was found to be 0.01% and obtained efficiency was 101.2%. Spiking of peanut powder in maize flour was performed at different concentrations.