Abstract: Microneedles (MNs) have emerged in the field of transdermal drug delivery due to their advantages of convenience, minimal invasiveness, and painlessness. In recent years, with the development of point-of-care testing (POCT), integrated microneedles in combination with sampling and real-time biomarker detection have received increasing attention in scientific research and medical service. This review summarizes the classification of microneedle-mediated diagnostic strategies, the types of samples that can be extracted by microneedles, and microneedle-mediated biomonitoring and diagnosis, with a prospect to the future development and application of microneedle-mediated biomonitoring and diagnosis.