The invention relates to a universal fluorescent primers based multiple competitive PCR method for detecting copy number variation. The method comprises the following steps: (1) providing multiple competitive PCR primers containing at least a pair of universal primers, at least a pair of chimeric specific primers located in a test section and at least a pair of chimeric specific primers located in a reference section; (2) constructing a full-length internal control sequence, and acquiring all the internal reference fragments through restriction enzyme digestion; (3) conducting a multiple competitive PCR reaction; and (4) analyzing data. The invention also provides a kit based on the above analysis method; and the method is suitable for detection of copy number variation of 5-28 genes / reactions, and is a fast, middle-flux and economical copy number variation detection scheme.