Genotyping with a dried blood spot method: a useful technique for application in pharmacogenetics.
Several commercial DNA isolation kits are available for extracting the genomic DNA from the ethylene diamine tetra-acetic acid (EDTA) whole blood samples. To obtain DNA from whole blood these DNA isolation procedures require quite some hands on time and are rather expensive. An alternative technique could be dried blood spot (DBS) sampling, with which DNA isolation is faster, cheaper and logistics are easier. We have developed a non-commercial DBS method and examined its performance in practice. DNA isolation from EDTA blood samples and made blood spots on filter paper from 106 renal transplant recipients were compared. Additionally, DNA isolation with a column method and two different DBS method was performed for 10 healthy volunteers and compared. Also DNA isolation with only capillary blood using both DBS methods from another 100 healthy volunteers has been investigated. Real-time PCR FRET assays for the CYP3A4 A-392G, CYP3A5 A6986G, ABCB1 C1236T, G2677T/A and C3435T polymorphisms were used and the melting curves of both DNA isolation methods were compared. In all cases DNA extracted with the column method corresponded completely with the results of the DNA isolated with the DBS procedure. Hence, DNA isolation from filter paper appears to be a useful alternative for the commercially available DNA isolation kits.
Wijnen
Petal A H M
PA
Department of Clinical Chemistry, University Hospital Maastricht, Maastricht, The Netherlands.
Op den Buijsch
Robert A M
RA
Cheung
Simon C Y
SC
van der Heijden
Jacques
J
Hoogtanders
Karin
K
Stolk
Leo M L
LM
van Dieijen-Visser
Marja P
MP
Neef
Cees
C
Drent
Marjolein
M
Bekers
Otto
O
D000483:Alleles; D001798:Blood Proteins / Q000235:genetics*; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D005821:Genetic Techniques; D005838:Genotype; D006801:Humans; D010597:Pharmacogenetics / Q000379:methods*; D011110:Polymorphism, Genetic / Q000235:genetics
D001798:Blood Proteins; D003577:Cytochrome P-450 Enzyme System; C510164:CYP3A5 protein, human; D051544:Cytochrome P-450 CYP3A