•Exosomes mediate cell-to-cell communication by transfer of functional substances such as nucleic acids, proteins, etc.•Exosomes contain many components which may represent biomarkers.•Exosomes are ideal candidates for non-invasive biomarker for several diseases.•Exosomes are considered the best biomarkers for cancer and several disease diagnosis. Exosomes are endosomal-derived vesicles that play a critical role in cell-to-cell communication and are secreted in several biological fluids including serum, saliva, urine, ascites, and cerebro-spinal fluid amongst others. Exosomes are small (30–150 nm diameter) with a distinctive bilipid protein structure. They can carry and exchange various cargos between cells and are used as a non-invasive biomarker for several diseases. Exosomes are considered the best biomarkers for cancer diagnosis, owing to their unique characteristics. Here, we provide a review of the up-to-date applications of exosomes, derived from various sources, in the prognosis and diagnosis of several diseases including cancer, cardiovascular and regenerative diseases as well as, arthritis, neurological diseases, and diabetes mellitus. The role of exosomes and their applications in biomedical research and preclinical trials have also been briefly discussed.