Feasibility and quality development of biomaterials in the pretest studies of the German National Cohort
Feasibility and quality development of biomaterials in the pretest studies of the German National Cohort
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DOI：
10.1007/s00103-014-2048-7
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发表时间：
2014-11-01
影响因子：
1.7
通讯作者：
Illig, Thomas
中科院分区：
文献类型：
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作者：
Kuehn, A.;Nieters, A.;Illig, Thomas
德国国家队列(GNC)旨在解决有关主要慢性疾病(例如癌症、糖尿病、传染病、过敏性疾病、神经和心血管疾病)的各种可能原因的研究问题，并确定风险因素和预后生物标记物，以早期诊断和预防这些疾病。收集生物材料，结合问卷调查和医疗检查中的大量信息，是研究的核心组成部分之一。在2011至2013年间对德国国家队列进行的两项预试研究中，从一定数量的参与者那里收集了一系列生物材料。10个研究中心参与了前测1，18个研究中心参与了前测2。制定并评估了标准操作程序(SOP)，以最大限度地减少生物样本采集过程中的分析前伪影。在前测研究中，对样品收集/准备的可行性[前测1(A)]以及生物标记物和蛋白质组分析的质量控制[前测1(B)、(C)]进行了不同方面的调查。此外，还根据共同标准测试了为具体项目招募研究参与者的情况、所有研究中心在规定时间内的检查程序以及生物样本的运输和分散储存(预试2)。这些分析将作为2014年开始的GNC主要研究中生物材料收集的基础。参与者从人群中随机选择(n=1000名受试者在预试1的10个研究地点招募)被要求捐献血液、尿液、唾液和粪便样本。此外，研究地点采集了鼻咽拭子，参与者在家中采集了鼻拭子。在前测2中，18个研究地点(n=599名受试者)收集了与前测1基本相同的生物材料，并进行了生物标志物分析以检验生物材料的质量。在前测1和2中，几乎所有受邀参与者的生物材料都是可行的，没有重大问题。受试者的平均有效率为95%。作为一个重要的结果，我们发现，例如，在抽血后，细胞组分应该在第一个小时内从血浆和血清组分中分离出来，在长达6小时的时间内没有显著变化，在4a时是所有分析的生物标志物的积分。此外，使用蛋白质组学方法对样品的质量控制表明，不同储存条件下的蛋白质没有明显的聚集。所有开发的SOP在经过一些适应和修改后，都被验证可用于主要研究。此外，还开发和测试了电子和纸质文档表，以记录收集的生物材料的时间戳、体积、冻结时间和等分编号。在所有参与研究的地点收集生物材料是可行的，没有重大问题。然而，在某些情况下，处理时间太长。为了避免样品采集中出现分析前的伪影，研究地点之间必须进行适当的标准化。为了实现这一目标，血液和尿液采集必须适应液体处理机器人的具体使用条件，GNC主要研究的所有参与研究中心都将提供这种机器人。严格遵守标准操作规程、对工作人员进行全面培训和准确的文件记录是获得高质量样品以供以后分析的必要条件。这样获得的生物材料为GNC传染病和其他常见复杂疾病的研究提供了宝贵的资源。
The German National Cohort (GNC) is designed to address research questions concerning a wide range of possible causes of major chronic diseases (e.g. cancer, diabetes, infectious, allergic, neurologic and cardiovascular diseases) as well as to identify risk factors and prognostic biomarkers for early diagnosis and prevention of these diseases. The collection of biomaterials in combination with extensive information from questionnaires and medical examinations represents one of the central study components.In two pretest studies of the German National Cohort conducted between 2011 and 2013, a range of biomaterials from a defined number of participants was collected. Ten study centres were involved in pretest 1 and 18 study centres were involved in pretest 2. Standard operation procedures (SOP) were developed and evaluated to minimize pre-analytical artefacts during biosample collection. Within the pretest studies different aspects concerning feasibility of sample collection/preparation [pretest 1 (a)] and quality control of biomarkers and proteome analyses were investigated [pretest 1 (b), (c)]. Additionally, recruitment of study participants for specific projects and examination procedures of all study centres in a defined time period according to common standards as well as transportation and decentralized storage of biological samples were tested (pretest 2). These analyses will serve as the basis for the biomaterial collection in the main study of the GNC starting in 2014.Participants, randomly chosen from the population (n = 1000 subjects recruited at ten study sites in pretest 1) were asked to donate blood, urine, saliva and stool samples. Additionally, nasal and oropharyngeal swabs were collected at the study sites and nasal swabs were collected by the participants at home. SOPs for sample collection, preparation, storage and transportation were developed and adopted for pretest 2. In pretest 2, 18 study sites (n = 599 subjects) collected biomaterials mostly identical to pretest 1. Biomarker analyses to test the quality of the biomaterials were performed.In pretest 1 and 2, it was feasible to collect all biomaterials from nearly all invited participants without major problems. The mean response rate of the subjects was 95 %. As one important result we found for example that after blood draw the cellular fraction should be separated from the plasma and serum fractions during the first hour with no significant variation for up to 6 h at 4 a"integral for all analysed biomarkers. Moreover, quality control of samples using a proteomics approach showed no significant clustering of proteins according to different storage conditions. All developed SOPs were validated for use in the main study after some adaptation and modification. Additionally, electronic and paper documentation sheets were developed and tested to record time stamps, volumes, freezing times, and aliquot numbers of the collected biomaterials.The collection of the biomaterials was feasible without major problems at all participating study sites. However, the processing times were in some cases too long. To avoid pre-analytical artefacts in sample collection, appropriate standardisation among the study sites is necessary. To achieve this, blood and urine collection will have to be adapted to specific conditions of usage of liquid handling robots, which will be available at all participating study centres in the main study of the GNC. Strict compliance with the SOPs, thorough training of the staff and accurate documentation are mandatory to obtain high sample quality for later analyses. The so obtained biomaterials represent a valuable resource for research on infectious and other common complex diseases in the GNC.