HLA-Typing using saliva as a non- invasive sampling method in the MAHAK hematopiotic stem cell registration center

The DNA extracted from saliva seems to be a more suitable alternative than peripheral blood for HLA-Typing by PCR- sequence-specific oligonucleotide (SSO) method.Its DNA yield is similar to that of peripheral blood, both in terms of quantity and quality. In addition the sampling is easy with little risk, is low cost and its acceptance by patients make this non invasive technique ideal for carrying out large scale studies with volunteers.In the study, 2ml of saliva sample were collected from 5000 participants in the MAHAK hematopiotic stem cell registration center; DNA was extracted by automatic equipments based on magnetic bead technique; PCR-SSO was done and then PCR product analyzed using automatic equipment with reverse hybridization technique. For evaluation of sample stability, 100 remaining of saliva samples were randomly selected from 5000 participants and stored at -20, 4, 25 â—¦c for one week (each sample in three divided parts). Then DNA was extracted and quality and quantity of DNA were assessed by nanophotometery and agarose gel electrophoresis.The result of HLA-Typing was successfully recorded with a repeat of less than 2% of the total. There were no significant differences in DNA concentration, purity and integrity between the stored samples at 3 different temperatures and the DNA extracted in the sampling day. The successful performance of 5000 saliva HLA-Typing and the stability of the DNA extracted from saliva stored under different temperature conditions indicate that these samples can be used as a suitable source of DNA extraction in large-scale studies.

Keywords:

HLA-Typing , SSO , hematopiotic stem cell