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Nowadays, one of the biggest concerns for ELISA is still how to improve the sensitivity of the assay, and the luciferase\u2013luciferin reaction system has been noticed as a new detection method with high sensitivity. In this study, a luciferin\u2013luciferase reaction system was used as the detection method for a sandwich ELISA system. It was shown that this new system led to an increase in the detection sensitivity of at least two times when compared with the traditional horseradish peroxidase (HRP) detection method. Lastly, the serum levels of the human extracellular matrix 1 protein of breast cancer patients were determined by the new system, which were overall similar to the HRP chemiluminescent system. 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