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J Vis Exp. 2015 Mar 13;(97). doi: 10.3791/52619.

Dried blood spots--preparing and processing for use in immunoassays and in molecular techniques.

Author information

1
Institute of Virology, National Reference Centre for Hepatitis C, Essen University Hospital, University of Duisburg-Essen.
2
Institute of Virology, National Reference Centre for Hepatitis C, Essen University Hospital, University of Duisburg-Essen; stefan.ross@uni-due.de.

Abstract

The idea of collecting blood on a paper card and subsequently using the dried blood spots (DBS) for diagnostic purposes originated a century ago. Since then, DBS testing for decades has remained predominantly focused on the diagnosis of infectious diseases especially in resource-limited settings or the systematic screening of newborns for inherited metabolic disorders and only recently have a variety of new and innovative DBS applications begun to emerge. For many years, pre-analytical variables were only inappropriately considered in the field of DBS testing and even today, with the exception of newborn screening, the entire pre-analytical phase, which comprises the preparation and processing of DBS for their final analysis has not been standardized. Given this background, a comprehensive step-by-step protocol, which covers al the essential phases, is proposed, i.e., collection of blood; preparation of blood spots; drying of blood spots; storage and transportation of DBS; elution of DBS, and finally analyses of DBS eluates. The effectiveness of this protocol was first evaluated with 1,762 coupled serum/DBS pairs for detecting markers of hepatitis B virus, hepatitis C virus, and human immunodeficiency virus infections on an automated analytical platform. In a second step, the protocol was utilized during a pilot study, which was conducted on active drug users in the German cities of Berlin and Essen.

PMID:
25867233
PMCID:
PMC4397000
DOI:
10.3791/52619
[Indexed for MEDLINE]
Free PMC Article

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