Background: Orthodontics is outstanding among the medical branches with a higher number of predisposing factors for cross-infection. Awareness of efficient sterilization techniques is considered to be a very important subject. Aim: To investigate the efficacy of various disinfection methods on orthodontic clamping tweezers contaminated with bacteria and Candida albicans. Materials and method: Staphylococci, Streptococci and Candida albicans cultivated from the sample of the whole saliva were taken from one patient after 3 months of braces placement. The broth was prepared of Streptococcus and Staphylococcus, and Candida albicans to obtain 106-107 CFU/ml. The peaks of 25 clamping tweezers were cut to get 50 pieces; the ends of these pieces were polished. Total 25 pieces of clamping tweezer peaks were divided into 5 groups with 5 pieces in each tube exposed to the broth of Streptococcus and Staphylococcus mixture for 5 minutes. The other 25 clamping tweezers pieces were also divided in the same manner and exposed to the broth of Candida albicans for 5 minutes. The 25 pieces of each group were subdivided into 5 groups, 4 of which were disinfected with Saniswiss biosanitizer I, desident cavicide, glutaraldehyde 2%, peracetic acid 0.25%, and the 5th group was autoclaved. Result: All the disinfection methods used showed no growth of staphylococci and streptococci. However, when the tweezers were contaminated with Candida albicans, disinfection with desident cavicide solution showed growth of candida on Sabouraud’s agar while the use of an autoclave, glutaraldehyde 2%, peracetic acid 0.25% and biosanitizer I, recorded no growth on Sabouraud’s agar. Conclusion: Autoclave, peracetic acid, glutaraldehyde and biosanitizer I can be used to effectively eliminate Streptococcus, Staphylococcus and Candida albicans.