1. The Application of Validation and Proficiency Testing Concepts From Current Clinical Genetic Diagnostics For the Implementation of New Genetic Technologies Genomics 2015 Kathleen S. Wilson, M.D. The University of Texas Southwestern Medical Center

2. Advent of new genetic technologies utilized for clinical diagnostics is rapid.

3. Implementation of these evolving technologies is challenging.

4. Next Generation Sequencing Multiple, fragmented sequence reads assembled based on overlapping areas https://en.wikipedia.org/wiki/DNA_sequencing

5. Whole Exome Sequencing https://en.wikipedia.org/wiki/Exome_sequencing#/media/File:Exome_Sequencing_Workflow_1a.png

6. College of American Pathologists/American College of Medical Genetics Cytogenetics Resource Committee

7. Fluorescence in situ hybridization (FISH)

8. College of American Pathologists/American College of Medical Genetics Cytogenetics Resource Committee Fluorescence in situ hybridization (FISH) Cytogenomic microarray analysis (CMA)

9. College of American Pathologists/American College of Medical Genetics Cytogenetics Resource Committee Fluorescence in situ hybridization (FISH) Cytogenomic microarray analysis (CMA) Relevance for the rapidly evolving sequencing technologies

10. Lessons learned from prior technology have relevance for the clinical genetic diagnostics of the future.

11. 1. Method not analyte based

12. UTSW FISH Probe Validation

13. 1.Metaphase probes a.Unique sequence 1.Localization 2.Analytical validation b.Repeat sequence/whole chromosome paints 1.Localization 2.Analytical validation 2.Interphase probes (unique sequence and repeat sequence only) a. Localization b. Analytical validation

14. Lessons learned from prior technology have relevance for the clinical genetic diagnostics of the future. 1.Method not analyte based Smear preparations Dropped cell suspensions from cultures Monolayer cultures Uncultured amniocytes Cytospin preparations Touch preparations FFPE (formalyn fixed parraffin embedded tissues) (CYJ/K/L) Urine

15. Lessons learned from prior technology have relevance for the clinical genetic diagnostics of the future. 1.Method not analyte based BCR/ABL1 RUNX1T1/RUNX1 PML/RARA Microdeletion cocktails ETC…

16. Lessons learned from prior technology have relevance for the clinical genetic diagnostics of the future. 1.Method not analyte based Dropped cell suspensions from cultures (CYF) Urine (Urovysion) (CYI) FFPE (formalyn fixed parraffin embedded tissues) (CYJ/K/L) HER2 (CYH)

17. Lessons learned from prior technology have relevance for the clinical genetic diagnostics of the future. 1.Method not analyte based Dropped cell suspensions from cultures (CYF) Urine (Urovysion) (CYI)

18. Cytogenomic Microarray Analysis

19. This was the fifteenth CAP/ACMG Survey mailing for whole genome copy number changes by microarray technology. This was the eleventh GRADED CYCGH Survey. The number of participants enrolled in this Survey has been 27, 34, 50, 63, 72, 80, 92, 96, 123, 124, 144, 147, 158, 161, and 170 in the first through fifteenth Surveys, respectively. CYCGH- 01 and CYCGH-02 reached participant consensus of greater than 80% and were graded.

20. Lessons learned from prior technology have relevance for the clinical genetic diagnostics of the future. 1. Method not analyte based 2. Vendor provided Quality Control measures as part of assay development and validation

21. Lessons learned from prior technology have relevance for the clinical genetic diagnostics of the future. 1. Method not analyte based 2. Vendor provided Quality Control measures as part of assay development and validation

22. Lessons learned from prior technology have relevance for the clinical genetic diagnostics of the future. 1. Method not analyte based 2. Vendor provided Quality Control measures as part of assay development and validation

23. Lessons learned from prior technology have relevance for the clinical genetic diagnostics of the future. 1. Method not analyte based 2. Vendor provided Quality Control measures as part of assay development and validation 3. DNA quality

24. Lessons learned from prior technology have relevance for the clinical genetic diagnostics of the future. 1. Method not analyte based 2. Vendor provided Quality Control measures as part of assay development and validation 3. DNA quality

25. Lessons learned from prior technology have relevance for the clinical genetic diagnostics of the future. 1. Method not analyte based 2. Vendor provided Quality Control measures as part of assay development and validation 3. DNA quality 4. Computer technology

26. Lessons learned from prior technology have relevance for the clinical genetic diagnostics of the future. 1. Method not analyte based 2. Vendor provided Quality Control measures as part of assay development and validation 3. DNA quality 4. Computer technology-interface checks

27. Lessons learned from prior technology have relevance for the clinical genetic diagnostics of the future. 1. Method not analyte based 2. Vendor provided Quality Control measures as part of assay development and validation 3. DNA quality 4. Computer technology-interface checks 5. Scope of validation