Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques
Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques
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DOI：
10.3791/52619
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发表时间：
2015-03-01
影响因子：
1.2
通讯作者：
Ross, R. Stefan
中科院分区：
文献类型：
--
作者：
Gruener, Nico;Stambouli, Oumaima;Ross, R. Stefan
在纸卡上收集血液，然后使用干燥的血斑（DBS）进行诊断的想法起源于一个世纪以前。从那时起，DBS测试几十年来一直主要集中在传染病的诊断上，特别是在资源有限的情况下或新生儿遗传代谢疾病的系统筛查上，直到最近才开始出现各种新的和创新的DBS应用。多年来，在DBS测试领域中，分析前变量只是被不恰当地考虑，即使在今天，除了新生儿筛查，整个分析前阶段，包括DBS的准备和处理，以进行最终分析，也没有标准化。鉴于这一背景，提出了一项涵盖所有基本阶段的全面分步方案，即采集血液；准备血斑；干燥血斑；星展银行的储存和运输；DBS的洗脱，最后对DBS洗脱液进行分析。该方案的有效性首先在一个自动分析平台上用1762对偶联血清/DBS对检测乙型肝炎病毒、丙型肝炎病毒和人类免疫缺陷病毒感染的标记物来评估。第二步，在对德国柏林和埃森两个城市的活跃吸毒者进行的一项试点研究中利用了该议定书。
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.