The invention belongs to the field of biotechnology detection, and particularly provides a Prader-Willi Syndrome detection kit and a use method thereof. According to the present invention, snO-lncRNA1, snO-lncRNA2, snO-lncRNA3, snO-lncRNA4 and snO-lncRNA5 are extremely highly expressed in human embryonic stem cells, and abnormally stable, so that the detection kit prepared by taking snO-lncRNA1, snO-lncRNA2, snO-lncRNA3, snO-lncRNA4 and snO-lncRNA5 as detection genes can detect samples such as common blood, saliva, urine and the like so as to provide guarantee for realizing non-invasive examination of Prader-Willi Syndrome; the use method is simple to operate and has low requirements on instruments and equipment; the input amount of the required sample is small, and the sample source is highly inclusive; the kit has the characteristics of high sensitivity, high specificity and the like; the detection result is visual, simple and easy to understand; and by using the detection method, Prader-Willi Syndrome can be detected in a short time.