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. 2014 Dec;5(6):351-7.
doi: 10.1016/j.phrp.2014.09.005. Epub 2014 Nov 1.

Rapid DNA extraction from dried blood spots on filter paper: potential applications in biobanking

Eun-Hye Choi  1 , Sang Kwang Lee  1 , Chunhwa Ihm  2 , Young-Hak Sohn  3
Affiliations

Rapid DNA extraction from dried blood spots on filter paper: potential applications in biobanking

Eun-Hye Choi et al. Osong Public Health Res Perspect. 2014 Dec.

Abstract

Objectives: Dried blood spot (DBS) technology is a microsampling alternative to traditional plasma or serum sampling for pharmaco- or toxicokinetic evaluation. DBS technology has been applied to diagnostic screening in drug discovery, nonclinical, and clinical settings. We have developed an improved elution protocol involving boiling of blood spots dried on Whatman filter paper.

Methods: The purpose of this study was to compare the quality, purity, and quantity of DNA isolated from frozen blood samples and DBSs. We optimized a method for extraction and estimation of DNA from blood spots dried on filter paper (3-mm FTA card). A single DBS containing 40 μL blood was used.

Results: DNA was efficiently extracted in phosphate-buffered saline (PBS) or Tris-EDTA (TE) buffer by incubation at 37°C overnight. DNA was stable in DBSs that were stored at room temperature or frozen. The housekeeping genes GAPDH and beta-actin were used as positive standards for polymerase chain reaction (PCR) validation of general diagnostic screening.

Conclusion: Our simple and convenient DBS storage and extraction methods are suitable for diagnostic screening by using very small volumes of blood collected on filter paper, and can be used in biobanks for blood sample storage.

Keywords: biobank; dried blood spot; filter paper.

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Figures

Figure 1
Figure 1
Workflow diagram. dsDNA = double-stranded DNA; RBC = red blood cell; ssDNA = single-stranded DNA.
Figure 2
Figure 2
DNA recovery from whole blood and Whatman FTA cards. BC = buffy coat in dried blood spots stored at room temperature; dsDNA = double-stranded DNA; WB = whole blood in dried blood spots stored at room temperature.
Figure 3
Figure 3
Average DNA yield over time. In the figure, fluid represents whole blood stored at −80°C and paper represents Whatman FTA cards stored at room temperature. ssDNA = single-stranded DNA.
Figure 4
Figure 4
DNA extracted by boiling dried blood spots on filter paper after incubation at 56°C. BC = buffy coat in dried blood spots stored at room temperature; ssDNA = single-stranded DNA; WB = whole blood in dried blood spots stored at room temperature.
Figure 5
Figure 5
Yield of ssDNA from dried blood spots stored at −80°C and room temperature. ssDNA = single-stranded DNA.
Figure 6
Figure 6
Efficiency of ssDNA extraction in PBS and TE. PBS = phosphate-buffered saline; ssDNA = single-stranded DNA.
Figure 7
Figure 7
PCR amplification of GAPDH and β-actin DNA. DNA was extracted from frozen-liquid samples and dried blood spots. Lanes 1–2 show dsDNA isolated from frozen whole blood and the buffy coat; lanes 3–4, dsDNA isolated from DBSs, with whole blood stored at room temperature; and lanes 5–6, ssDNA isolated from DBSs, with whole blood and the buffy coat stored at room temperature and extracted with the boiling method. BC = buffy coat in dried blood spots stored at room temperature; DBS = dried blood spot; dsDNA = double-stranded DNA; PCR = polymerase chain reaction; ssDNA = single-stranded DNA; WB = whole blood in dried blood spots stored at room temperature.
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References

    1. Petrini C., Olivieri A., Corbetta C. Common criteria among States for storage and use of dried blood spot specimens after newborn screening. Ann Ist Super Sanita. 2012;48(2):119–121. - PubMed
    1. Lang P.O., Govind S., Drame M. Comparison of manual and automated DNA purification for measuring TREC in dried blood spot (DBS) samples with qPCR. J Immunol Methods. 2012 Oct;384(1–2):118–127. - PubMed
    1. Guthrie R., Susi A. A simple phenylalanine method for detecting phenylketonuria in large populations of newborn infants. Pediatrics. 1963 Sep;32:338–343. - PubMed
    1. Lindstrom B., Ericsson O., Alvan G. Determination of chloroquine and its desethyl metabolite in whole blood: an application for samples collected in capillary tubes and dried on filter paper. Ther Drug Monit. 1985;7(2):207–210. - PubMed
    1. Prior T.W., Highsmith W.E., Jr., Friedman K.J. A model for molecular screening of newborns: simultaneous detection of Duchenne/Becker muscular dystrophies and cystic fibrosis. Clin Chem. 1990 Oct;36(10):1756–1759. - PubMed