Abstract: Aim: Collection of cheek buccal cells using swabs provides a reliable method for obtaining DNA samples from donors who join the Canadian OneMatch Stem Cell and Marrow Network. In this study, we evaluated the impact of storage temperature [room temperature (RT), -30°C and -80°C] during a five-year period on obtaining accurate HLA-A, -B, DRB1 typing. Methods: Buccal swabs were collected from six previously typed volunteers, stored at room and cold temperatures and tested at one-year intervals for a period of five years. At the end of each year of storage, samples were extracted, gel electrophoresis on amplified products was performed to assess their integrity and quality, and HLA typing was performed using the LABType rSSO, Micro SSP (One Lambda, Inc.) and AlleleSEQR® SBT (Atria Genetics, Abbott) kits. Results: Amplified DNA from all samples stored at either RT or cold temperatures showed no degradation and expected amplified products by gel electrophoresis and accurate typing results were obtained for up to five years using SSO and SSP methods. Using SBT method, we observed that, compared to cold storage, samples stored at RT had overall lower electropherogram signal intensity. From year two to five of storage, the average range in reduction of signal intensity was 82 - 94% for HLA-A, 47 - 92% for HLA-B and 6 - 43% for DRB1. Accurate HLA typing was obtained by SBT for HLA-DRB1 for all samples stored up to five years at either cold or RT. The SBT HLA typing for HLA-A was 100% successful up to year 2 of storage at RT, dropped to 67% and 17% at years 3 and 4, respectively, and failed 100% at year 5. The success rate for HLA-B by SBT was 100% for up to year 3 and was reduced to 83% and 33% at years 4 and 5, respectively. Conclusions: In summary, the buccal swab method is a robust, cost effective way to collect DNA for HLA typing of unrelated donors. The HLA typing success rate for cold storage was 100% for all three methods up to five years. Compared to cold storage, RT storage of buccal swab samples up to five years gave accurate typing results for all DNA samples tested by SSO and SSP for all three loci and by SBT for HLA- DRB1. No typing could be obtained for HLA-A by SBT by year 5 of RT storage. [Copyright &y& Elsevier]