VWF Phenotyping and Molecular Analysis Core

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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.