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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

12 Issues per year


IMPACT FACTOR 2016: 3.432

CiteScore 2016: 2.21

SCImago Journal Rank (SJR) 2016: 1.000
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1437-4331
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Volume 47, Issue 12 (Dec 2009)

Issues

Quantitation of RNA decay in dried blood spots during 20 years of storage

Fredrika Gauffin
  • Division of Pediatrics, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet, Stockholm, Sweden
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Ann Nordgren
  • Division of Clinical Genetics, Department of Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Gisela Barbany
  • Division of Clinical Genetics, Department of Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Britt Gustafsson
  • Division of Pediatrics, Department of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet, Stockholm, Sweden
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Håkan Karlsson
Published Online: 2009-10-28 | DOI: https://doi.org/10.1515/CCLM.2009.351

Abstract

Background: Diseases with an onset during childhood or adult life can have their origin during fetal life or at birth. Neonatal blood dried on filter paper (Guthrie cards) collected for screening purposes is routinely stored for decades. In addition to clinical use, these filters in combination with patient registers constitute an invaluable resource for epidemiological and pathophysiological research. Although RNA has been successfully recovered from such filters even after decades of storage, the potential decay of RNA over time has not previously been investigated using quantitative methods.

Methods: Filter papers (n=5) with dried blood spots from the Swedish National PKU register, stored for 1, 5, 10, 15 or 20 years were randomly selected. RNA was isolated from each sample, quantitated by spectrophotometry and reverse transcribed following DNase I treatment. Amplifiable cDNA was subsequently detected by real-time PCR using primers specific for transcripts encoding β-actin.

Results: Transcripts encoding β-actin were detected in all 25 samples analyzed at a mean threshold cycle (Ct) of 25 (SD 1.9). A one-way ANOVA indicated no significant effect of storage time on Ct values.

Conclusions: The lack of significant decay of RNA in dried blood filters stored for up to 20 years suggests that such filters are useful for studies of RNA determinants of diseases with an onset in childhood as well as adult life.

Clin Chem Lab Med 2009;47:1467–9.

Keywords: dried blood spots; Guthrie cards; quantitative real-time PCR; RNA

About the article

Corresponding author: Håkan Karlsson, Department of Neuroscience, Karolinska Institutet, Retzius väg 8, 171 77 Stockholm, Sweden Phone: +46 8 524 878 32, Fax: +46 8 325 325,


Received: 2009-07-03

Accepted: 2009-09-01

Published Online: 2009-10-28

Published in Print: 2009-12-01


Citation Information: Clinical Chemistry and Laboratory Medicine, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2009.351.

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©2009 by Walter de Gruyter Berlin New York. Copyright Clearance Center

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