Background and Objective: Many polymorphisms had been mapped in vitamin D receptor (VDR) gene on chromosome 12, they had variable location which influences its functional prospect. VDR FokI single nucleotide polymorphism (SNP) that resulted from T/C substitution at exon 2 leads to alteration of the first transcription site. The present study aimed to evaluate the possible association between VDR gene FokI SNP (C > T, rs2228570) polymorphism and spontaneous preterm birth (SPTB) in the Egyptian pregnant population. Materials and Methods: A total of 160 pregnant women, divided into two groups: 80 preterm (case group) and 80 control group, were investigated for VDR gene FokI polymorphism by predesigned Taq-Man SNPs genotyping assay. Results: The odds ratio (OR) for the preterm birth risk was significantly higher with the mutant homozygous (TT) genotype (p = 0.01). Vitamin D level is significantly different among the three VDR FokI SNP genotypes in both groups. The lowest vitamin D level was found in variant homozygous TT genotype (19.6 and 25.5 ng mL-1 in preterm and control groups, respectively). The C allele was associated with low preterm birth risk [OR: 0.43 (0.20- 0.91)], and the T allele was associated with high preterm labor risk [OR: 2.0 (1.04-3.79)]. Conclusion: VDR FokI polymorphism is associated with an increased risk of spontaneous preterm birth (SPTB) in Egyptian pregnant women. It may play a possible role in SPTB etiology. The current study provides data on the consideration of VDR FokI polymorphism as a potential biomarker for SPTB.