Since 2010, matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS) has been used in healthcare settings. Over the traditional method of biochemical identification, MALDI-TOF MS has a number of advantages, including simplicity, speed, precision, and affordability. Numerous challenges to detecting bacteria, fungi, and viruses can be overcome using this method. As technology developed, a growing number of databases tracked spectra, enabling the identification of species that had similar morphological, genotypic, and biochemical features. Due to improvements in sample preparation and database enrichment, using MALDI-TOF MS for identification has become more precise and rapid. MALDI-TOF MS has yielded encouraging results for colony identification and quick sample detection. Rapidly diagnosing extremely contagious and drug- resistant illnesses is a crucial use of MALDI-TOF MS. Here, we give a review of the scientific literature evaluating the efficiency of MALDI-TOF MS for identifying pathogenic bacteria, fungi, and viruses that are relevant to clinical settings. Although MALDI-TOF MS offers several advantages for finding clinical infections, it also has significant disadvantages. Because there aren't many spectra in the database and organisms have inherent similarities, it might be challenging to tell one species from another, leading to misidentifications. Additional testing is typically necessary to fix these issues, which are extremely rare. In conclusion, the use of MALDI-TOF is revolutionising the detection and treatment of infectious diseases.