To develop a specific method for the detection of S. aureus , chicken anti-protein A IgY was adopted for specifically capturing S. aureus , depending on the specific recognition of staphylococcal protein A (SPA) by chicken anti-protein A IgY, which can eliminate the interference from protein G-producing Streptococcus. HRP labeled IgG, Fc region of which has a high affinity towards SPA, was paired with IgY for the colorimeter analysis of the system. By optimizing the system, a super-low detection limit of 11 CFU of S. aureus in 100 μL PBS without enrichment, with a linear range from 5.0 × 102 CFU mL−1 to 5.0 × 104 CFU mL−1 was obtained. The entire assay was accomplished in less than 90 min and no cross-reactivity with the other tested bacterial species was observed. Moreover, the developed assay has been applied for the detection of S. aureus in three different types of real samples (sodium chloride injection, apple juice and human urine) with satisfactory results. To the best of our knowledge, it is the first time to report using chicken anti-protein A IgY and any IgG to detect S. aureus based on the dual-recognition mode of SPA. The novel method opened up a way for monitoring S. aureus in food samples with high sensitivity, specificity and simple operation. • Chicken anti-protein A IgY was utilized to capture S. aureus with high selectivity. • HRP-IgG, with high affinity to SPA of S. aureus , was adopted for colorimetry. • 11 CFU of S. aureus in 100 μL samples were detected with high specificity. • No interference from protein G producing Streptococcus was observed. • The method has been applied to detect S. aureus in various types of samples. [ABSTRACT FROM AUTHOR]