Relating to the quarantine inspection field of plant diseases and insect pests, the invention provides a Pratylenchus penetrans detection kit and a detection method thereof. By designing specific primers specific to the 18S specific sequence of Pratylenchus penetrans and applying a PCR amplification technology, rapid detection of Pratylenchus penetrans is realized. With the characteristics of strong specificity, high sensitivity and good stability, the method plays an important role in improving the accuracy of Pratylenchus penetrans detection and shortening the detection time.