The invention relates to the field of temperature calibration of polymerase chain reaction analyzers, in particular to a multi-channel temperature calibration device of a polymerase chain reaction analyzer, which comprises a plurality of temperature sensors. The signal output ends of the plurality of temperature sensors are connected with a multi-channel change over switch, the multi-channel change over switch is connected with an integrated temperature measurement module, the integrated temperature measurement module is connected with a micro processing module, and the output end of the microprocessing module is connected with the control end of the multi-channel change over switch. The micro processing module is connected with a computer control unit which is used for displaying, storing information and generating a calibration report; each temperature sensor comprises 15 plug-in thermistor sensors and one patch thermistor sensor; the micro processing module controls the multi-channel change cover switch through a protocol signal control circuit; and the integrated temperature measurement module is a comparative temperature measurement circuit.