Abstract: Luminex assay belongs to a high throughput biochip detecting platform, which is based on the combination of fluorescence-coded microspheres, laser test, flow cytometry and data analysis system. This technique enables simultaneous detection of as many as 100, even 500 kinds of different target molecules, including proteins, peptides, DNA and RNA. It is well accepted for the advantages, such as good reproducibility, high sensitivity, strong specificity and easy operation. The aim of the review is to summarize the mechanism of Luminex assay, system composition, various biological applications and comparison with other protein and nucleic acid testing techniques. Luminex assay will have broad prospects in the research of pure science and clinical trials.