Traditionally, in Australia, blood samples have been collected from unrelated bone marrow donor volunteers for recruitment to the Australian Bone Marrow Donor Registry (ABMDR), while larger worldwide registries have predominantly utilised dry buccal swabs. Dry buccal swabs provide the advantage as they are less invasive, do not require a phlebotomist, are cost efficient and provide an easier solution for distribution to volunteers which can potentially increase donor participation. Dry buccal swabs are also more suitable to obtain DNA from difficult to bleed patients, young children and patients with low cell counts in which HLA typing or molecular based assays are requested.
We evaluated COPAN dry swabs in conjunction with the Maxwell™ HT 96 gDNA Blood Kit to determine suitable gDNA quantity and quality for downstream molecular applications such as HLA typing and ABO blood grouping using third generation sequencing technology.
Twenty one participants consented to providing dry buccal swabs for this evaluation. Participants were provided instructions for buccal collection based on manufacturer instructions. The buccal swabs were extracted using Maxwell HT 96 gDNA Blood kit on the fully automated King Fisher FLEX system by Thermofisher.
All dry buccal swabs processed, resulted in DNA yields >25ng/ul and acceptable 260/280 ratio’s. Degradation of DNA, evident by a smear during gel electrophoresis, was observed in five of the twenty two samples. This is expected as bacteria and nucleases present in unprocessed buccal swabs will cause DNA degradation. It is therefore recommended to increase the starting material of buccal DNA in downstream applications.
Dry buccal swab collection is a non-invasive DNA collection method suitable for use in replacement of blood . Isolation of DNA using the COPAN FLOQ dry buccal swabs in conjunction with the Promega Maxwell™ HT 96 gDNA Blood Kit provides suitable gDNA for use in downstream molecular based methods.