This study was purposed to investigate the relationship between NQO1C(609T), RAD51(G135C), XRCC3(C241T) single nucleotide polymorphisms and incidence of acute lymphoblastic leukemia (ALL). NQO1C(609T), RAD51(G135C), XRCC3(C241T) genotypes were detected by PCR-RFLP in 170 patients with de novo ALL and 458 normal persons as control. The results indicated that the genotype ratio of NQO1C(609T), RAD51(G135C) and XRCC3(C241T) in single genotype analysis showed no statistical difference between ALL patients and normal controls, which suggested that the single genotype affect onset of ALL without statistical significance. In combined genotype analysis, presence of both variants for NQO1C(609T) and RAD51(G135C) increased onset risk of ALL with myeloid antigen positive and with balanced translocation (OR value 5.553 and 2.618 respectively); the presence of homozygosity variant for NQO1C(609T) increased onset risk of ALL in the country-children (OR = 2.541). In conclusion, the combined effect of NQO1C(609T), RAD51(G135C) and XRCC3(C241T) genotypes may promote occurrence of ALL, which suggests that the combined analysis of 3 genotypes has more predictive significance for ALL than single genotype analysis.