The invention provides a diabetic retinopathy related gene polymorphism rapid detection kit. The kit comprises multiplex PCR pre-amplification primers and single base extension primer sequences of 23 single nucleotide polymorphism (SNP) sites relates to diabetic retinopathy, a human peripheral blood genome DNA extraction liquid, multiplex PCR pre-amplification reaction liquid, a negative control, a positive control, a pre-amplification product purification reagent, PCR product single base extension reaction solution liquid, and a extension product purification reagent. The invention also provided a preparation method of the kit. According to the invention, diabetic retinopathy related SNP sites are detected by using a gene sequencing method with relatively high sensitivity and specificity, such that genotypes of the 23 diabetic retinopathy related gene polymorphism sites can be detected rapidly, effectively, accurately, and economically. Therefore, a reference is provided for diabetic retinopathy early diagnosis and clinical prognosis.