The Luminex test can detect low levels of donor-specific antibody (DSA) that could not be detected by flow-cytometric crossmatching (FCXM) in kidney transplantation (KT). This study evaluated the impact of DSA on clinical outcomes in KT recipients negative on FCXM. Of 575 consecutive patients who underwent living donor KT between January 2013 and July 2016, 494 (85.9%) were DSA-negative and 81 (14.1%) were DSA-positive. Although rates of acute cellular rejection (ACR) at 1 year were similar in the two groups (P = 0.54), the incidence of antibody-mediated rejection (ABMR) was significantly higher in the DSA-positive group (P < 0.01). There was no statistically significant association between rejection-free graft survival (RFGS) rates and pretransplant class I DSA. However, evaluation of pretransplant class II DSA showed that RFGS rates were significantly lower in patients with MFI >3000 than DSA negative (P < 0.01). On multivariate analyses, class II DSA MFI ≥5000 was a significant risk factor for acute rejection (hazard ratio, 7.48; P < 0.01). These findings suggested that pretransplant DSA alone did not affect graft survival in KT recipients without desensitization. However, class II DSA MFI >5000 was an independent predictor of acute rejection in patients DSA positive. Patients at high risk of AR will therefore require pretransplant desensitization treatment or a strong induction agent to prevent early rejection after KT.