A cartridge for digital real-time Polymerase chain reaction (PCR) includes a microfluidic chamber, a well array, a CMOS photo sensor array and a PCB. The microfluidic chamber includes an inlet formed for injection of a liquid sample, the microfluidic chamber being capable of injection molding. The well array includes a plurality of microwells through which upper and lower portions are perforated and being attached to a lower surface of the microfluidic chamber. The CMOS photo sensor array is disposed below the well array to capture a response image of a sample filled in microwells of the well array. The PCB has a vent formed for vacuum processing of micro flow path formed in the microfluidic chamber, a space formed between the well array and the microfluidic chamber, and a microwell formed in the well array as the liquid sample is injected through the inlet.