VWF Phenotyping and Molecular Analysis Core

项目总结（见说明）： 核心B在Dan Bellissimo博士的指导下，由Ken Friedman博士提供重要的额外投入。 该核心具有许多重要功能，包括1）样品接受和中央处理; 2)添加血液中心跟踪标签，用于BCW诊断实验室的注册目的。这些样本上的去识别标签由各个临床中心放置。VWF表型的样本检测在Friedman博士指导的止血实验室进行。它们不是按照设计进行批处理的，因此这些样本的结果是以与患者样本提交实验室相同的方式获得的。当结果完成、审核并签出时，结果将自动传输至Investig 8数据库。DNA测序有两种类型。第一种是直接的，基于PCR的VWF基因的编码区测序，然后进行毛细管凝胶分析。在前一周期的第4年，该职能从波士顿转移至密尔沃基。不仅为临床实验室中的所有检测设置了内标对照品，而且核心A具有正常、临界、中度和中重度VWD血浆的重复等分试样，用于确保PPG研究的QG。新一代测序已被添加到该核心功能中，并将用于项目1和3。 虽然下一代测序的成本和设备需求正在迅速下降，但这包括在Corte B中，以便我们可以集中指导PPG所需的质量和成本。我们预计在北美有一个中心（多伦多大学或威斯康星州医学院），在英国曼彻斯特有一个。这两个站点必须仔细交叉标准化，以便数据输出相同。在欧盟内设立第二个研究中心的原因是MCMDM-1 VWD为这些样本开发的系统无法使样本离开参与合作伙伴的国家。

PROJECT SUMMARY (See instructions): Core B is under the direction of Dan Bellissimo, PhD with important additional input by Ken Friedman, MD. This core serves a number of important functions including 1) sample acceptance and central processing; 2) adding blood center tracking labels for registration purposes of the BCW Diagnostics Laboratory. The de-identified labels on these samples were put in place by the individual clinical centers. The sample testing for VWF phenotyping is done in the Hemostasis Laboratory directed by Dr. Friedman. They are NOT batched by design so that the results obtained on these samples are obtained in the same manner as if a patient sample was referred to the laboratory. When results are completed, reviewed, and signed out the results are automatically transferred to the Investig8 Database. DNA sequencing is of two types. The first is direct, PCR-based sequencing of the coding region of the VWF gene followed by capillary gel analysis. This function was transferred to Milwaukee from Boston during year 4 of the prior cycle. Not only are there the internal standard controls set in place for all testing in the clinical labs, but Core A has repetitive aliquots of normal, borderline, moderate and moderately severe VWD plasma that are used to assure the QG for the PPG study. Next Gen sequencing has been added to this core function and will be used in Projects 1 and 3. While the cost and equipment needs for Next Gen sequencing are rapidly coming down, this is included in Corte B so that we can centrally direct the quality and costs that will be required by the PPG. We anticipate one center in North America (either University of Toronto or Medical College of Wisconsin) and one in Manchester, UK. These two site must be carefully cross standardized so that data output is identical. The reason for the second site within the EU is because of the system developed for these samples by the MCMDM-1 VWD that does not enable samples to leave the Countries of the participating partners.