Rough Set theory is the mathematic theory that can express and deal with vague and uncertain data.There is complicated and uncertain data in the fault feature of Nuclear Power Plant,so that Rough Set theory can be introduced to analyze and process the historical data to find out the rule of fault diagnosis of Nuclear Power Plant.This paper introduces the Rough Set theory and Knowledge Acquisition briefly,and describes the reduction algorithm based on discernibility matrix and its application in the fault diagnosis to generate rules of diagnosis.Using these rules,three kinds of model faults have been diagnosed correctly.The conclusion can be drawn that this method can reduce the redundancy of the fault feature,simplify and optimize the rule of diagnosis.