The invention relates to a two-path temperature control polymerase chain reactor and a real-time detection device, which are applied in the field of biochemical reaction and medical detection. The two-path temperature control polymerase chain reactor comprises a base layer with a ring-shaped microfluidic channel structure, wherein the base layer is sealed with a cover board layer to form a microfluidic chip, the cover board layer is provided with a sample incoming hole and a sample outgoing hole, a microprocessor is adopted to control two semi-conductor heating and refrigerating devices, a whole microfluidic channel area forms three constant temperature areas, namely a degeneration area, an annealing area and an extension area needed by polymerase chain reaction, the semi-conductor heating and refrigerating devices are arranged under the microfluidic chip, and a platinum resistance temperature sensor manufactured by a semi-conductor process is arranged among the microfluidic chip and the two semi-conductor heating and refrigerating devices, so that temperature feedback signals are provided to the microprocessor, and closed ring control can be formed. The real-time detection systemcomprises a light source, a photoelectric detector and a fluorescence signal acquisition processing system. In the invention, by controlling the two semi-conductor heating and refrigerating devices, the whole microfluidic channel area forms the three constant temperature areas, and the heating and refrigerating device is simplified.