Introduction The need for accurate relative biological effectiveness (RBE) estimation for low energy therapeutic X-rays (corresponding to 50 kV nominal energy of a commercial low-energy IORT system (INTRABEAM)) is a crucial issue due to increased radiobiological effects, respect to high energy photons. Modeling of radiation-induced DNA damage through Monte Carlo (MC) simulation approaches can give useful information. Hence, this study aimed to evaluate and compare RBE of low energy therapeutic X-rays using Geant4-DNA toolkit and Monte Carlo damage simulation (MCDS) code. Materials and methods RBE calculations were performed considering the emitted secondary electron spectra through interactions of low energy X-rays inside the medium. In Geant4-DNA, the DNA strand breaks were obtained by employing a B-DNA model in physical stage with 10.79 eV energy-threshold and the probability of hydroxyl radical’s chemical reactions of about 0.13%. Furthermore, RBE estimations by MCDS code were performed under fully aerobic conditions. Results Acquired results by two considered MC codes showed that the same trend is found for RBEDSB and RBESSB variations. Totally, a reasonable agreement between the calculated RBE values (both RBESSB and RBEDSB) existed between the two considered MC codes. The mean differences of 9.2% and 1.8% were obtained between the estimated RBESSB and RBEDSB values by two codes, respectively. Conclusion Based on the obtained results, it can be concluded that a tolerable accordance is found between the calculated RBEDSB values through MCDS and Geant4-DNA, a fact which appropriates both codes for RBE evaluations of low energy therapeutic X-rays, especially in the case of RBEDSB where lethal damages are regarded.