Point-of-care testing (POCT) is necessary to provide a rapid diagnostic result for a prompt on-site diagnosis and treatment. A quick analysis time and high sensitivity, with a sample-to-answer format, are the most important features for current POCT diagnostic systems. Microfluidic lab-on-a-chip technologies have been considered as one of the promising solutions that can meet the requirement of the POCT since they can miniaturize and integrate most of the functional modules used in central laboratories into a small chip. This review covers recent advances in POCT technologies with an emphasis on demonstrated and commercially available POCT diagnostic systems with laboratory quality using microfluidic lab-on-a-chip technologies. As working principles and required functional modules depend on target analytes, we categorize the applications of the POCT diagnostic systems according to the analyte types such as proteins, cells, nucleic acids, and metabolites. In each analyte category, detection methods, configuration of POCT lab-on-a-chip modules, and advantages and disadvantages of POCT systems are reviewed and discussed along with future prospects.