title: Comparison of manual and automated DNA purification for measuring TREC in dried blood spot (DBS) samples with qPCR.
abstract: Automated nucleic acid extractions from dried blood spot (DBS) samples promises standardized sample treatment, low error rates, avoidance of contamination and requirement of less hands-on time. In the present study, non-automated and automated column based extraction processes using the QIAamp Investigator procedure were compared for the extraction of DNA from DBS samples. The concentration and the purity of DNA generated were determined by optical density readings. Furthermore qPCR downstream applications using the nucleic acids extracted with the two processes and albumin and T-cell receptor excision circles (TREC) copy numbers were measured and compared. The influence of the time of storage was also investigated by analyzing samples freshly dried and stored up to 11weeks at -20°C from the same individual. Finally, we provide arguments of preferentially choosing the automated procedure for extracting DNAs from DBS samples when downstream qPCR applications are required.
authors.affiliation: Translational Medicine Research Group, Cranfield Health, Cranfield University, Cranfield, United Kingdom. polang@nescens.com
authors.forename: Pierre-Olivier
authors.initials: PO
authors.lastname: Lang
authors.forename: Sheila
authors.initials: S
authors.lastname: Govind
authors.forename: Moustapha
authors.initials: M
authors.lastname: Dramé
authors.forename: Richard
authors.initials: R
authors.lastname: Aspinall
mesh_terms: D001709:Biotechnology / Q000295:instrumentation / Q000379:methods; D015925:Cryopreservation; D004247:DNA / Q000097:blood / Q000235:genetics / Q000302:isolation & purification*; D059788:Dried Blood Spot Testing / Q000295:instrumentation / Q000379:methods*; D015329:Gene Rearrangement, T-Lymphocyte; D006801:Humans; D007963:Leukocytes, Mononuclear; D016133:Polymerase Chain Reaction / Q000379:methods; D011933:Reagent Kits, Diagnostic; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000235:genetics; D013997:Time Factors
chemical_list: D011933:Reagent Kits, Diagnostic; D011948:Receptors, Antigen, T-Cell; D012709:Serum Albumin; D004247:DNA