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We investigated the genetic markers associated with elevated serum prostate-specific antigen (sPSA) levels to improve the predictive power of sPSA in screening for prostate cancer. A genome-wide association study was carried out among 4124 healthy Korean male adults using the Affymetrix Axiom Customized Biobank Genotyping Arrays for sPSA levels. A subgroup analysis for increased sPSA levels who underwent a prostate biopsy (n=64) was also carried out. We detected 11 single nucleotide polymorphisms (SNPs) near the Solute carrier family 45member 3, AGAP7P, MSMB, LOC101929917, and KLK3 genes associated with sPSA levels. The top SNP associated with the log of the sPSA levels was rs72434280 in the Solute carrier family 45 member 3 gene (P value, discovery set=2.98×10−20, replication set=7.31×10−10). A case–control study utilizing available biopsy reports (49 patients with normal biopsies vs. 15 patients with biopsies indicating cancer) for the sPSA more than 3 ng/ml group was carried out for the respective SNPs after adjusting for age. Only the SNPs near the KLK3 gene were associated with prostate cancer. In the model of the predictive elevation of sPSA level, adding the genetic risk score [area under the curve (AUC)=0.697] to age and BMI (AUC=0.602) significantly improved the results of the AUC (P<0.0001). We found seven SNPs associated with elevated prostate-specific antigen levels in healthy Korean men. Four SNPs were a novel marker in the Korean population. In men with increased prostate-specific antigen levels, genotyping SNP related to cancer-free elevation of sPSA level could be informative to decide the indication of prostate biopsy.