Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques

被引:128
作者
Gruener, Nico [1 ]
Stambouli, Oumaima [1 ]
Ross, R. Stefan [1 ]
机构
[1] Univ Duisburg Essen, Essen Univ Hosp, Natl Reference Ctr Hepatitis C, Inst Virol, Essen, Germany
来源
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2015年 / 97期
关键词
Molecular Biology; Issue; 97; Dried blood spots; filter paper cards; specimen storage; infectious diseases; hepatitis B virus; hepatitis C virus; human immunodeficiency virus; HEPATITIS-C VIRUS; FILTER-PAPER; B-VIRUS; ANTIBODY DETECTION; TOOL; PERFORMANCE; INFECTIONS; COLLECTION; ANTIGEN; SAMPLES;
D O I
10.3791/52619
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
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页数:9
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