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The successful treatment of most cancers depends on early detection. Tumor mRNA as a specific marker provides new avenues to monitor tumor progression in the early stages and assesses response to treatment. However, single tumor mRNA testing usually yields “false positive” results because cancer is associated with multiple tumor mRNA. It is indispensable to develop simple and effective approaches for the detection of multiple tumor mRNA. In this study, we used a combination of tumor‐specific mRNA markers to avoid the inherent limitations associated with the single‐marker technique. A gold nanoparticle (AuNP) was assembled with a bi‐molecular beacon (bi‐MB), and termed AuNP/bi‐MB, which simultaneously targeted to two types of tumor mRNA in breast cancer cells. This imaging agent could prevent effectively false positive results and provide comprehensive and dependable information for the …