What s in your DNA? Name: Date / Time: Event:

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1 What s in your DNA? Name: Sunitha Vege, MS Manager, Genomics Laboratory, NYBC Date / Time: Monday, October 8, 2012 / 7:45 AM Event: AABB Boston Industry Workshop 26 GLOBCC 5OCT2014

2 Background 1850 s: G. Mendel inheritance of traits; father of genetics 1950 s: Structure of DNA was discovered J. Watson and F. Crick R. Franklin and M. Wilkins 1980 s: Advent of PCR technology K. Mullis 2 What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only

3 Age of DNA-based Testing Biology Forensics Paternity Disease diagnosis Microbiology Pathology 3 What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only

4 Evolution of genomics: DNA Extraction Manual Semi-automated* Automated* 1-2 hours 1-2 hours 3 hours 12 sample 12 samples 96 samples *QIAcube and MDx instruments, QIAGEN, Valencia, CA 4 What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only

5 Evolution of genomics: PCR testing Manual assays Real-time PCR* Array technologies DNA probes on: colored beads Luminex *Eppendorf miniture beads on silicone chip Immucor/BioArray glass slides Days/Week Hours/Days hydrostatic holes <8 hours 5 What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only

6 Advantage of array technologies Type for multiple minor antigens in a single assay * BLOODchip Luminex Assays * * *In the US and Canada, the BLOODChip products are for research use only. Not for use in diagnostic procedures. 6 What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only

7 Current DNA-based testing for patients Multiply transfused: DNA isolated from white cells +DAT Allo versus auto antibodies OB patients to determine if candidate for Rh immune globulin RHD screen (weak/partial) to confirm RhD status - Currently done when serology typing is weaker than expected - Future: Test all OB patients Determine if candidate for Rh immune globulin Zygosity testing of paternal sample when mother has antibody, typically to RhD, -K, -c, or HPA-1a - Type fetus from amniocytes to determine if possibility of hemolytic disease newborn (HDN) or neonatal alloimmune thrombocytopenia (NAIT). 7 What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only

8 Testing of patients with Sickle Cell Disease (SCD) Full RH genotype - RH common - common Rh antigens are: D ( Rh positive or Rh negative ) C and c, E and e antigens Exceptions in African Black ethnic group - Many additional Rh antigens (n=>50 antigens) - Not detected with serologic reagents - Have significance for transfusion - Matching program for C, E, and K Not truly Rh antigen matched unless genotyped 8 What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only

9 Current DNA-based testing for donors Resolve typing discrepancies such as with Fyb, ABO, and Rh Genotype for antigens for which serologic reagents are not available or are not reliable Screen donors for rare phenotypes RH genotyping of donors, particularly of African Black ethnicity, for potential match for sickle cell patients with unexpected Rh antibodies 9 What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only

10 Molecular characterization of panel cells Currently, some commercial panel cells (Immucor) have extended antigen results listed by DNA testing, i.e. Dombrock In specialized reference laboratories, panel cells characterized for rare RH genotypes for resolving difficult Rh antibodies DIIIa ces / DIIIa ces r S / r S; i.e. (C)ceS DAR cear / DAR cear 10 What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only

11 Frequently asked questions Patient/donor consent: not genetic testing Determination of blood group antigens by different method Is a predicted phenotype Reimbursement for testing: CPT codes for molecular testing apply Standards: AABB 11 What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only

12 Frequently asked questions Not yet FDA approved research use only similar to unlicensed typing sera used in reference lab for typing uncommon antigens NYBC disclaimer These in vitro diagnostic tests were developed and their performance characteristics established in the Molecular Analysis Laboratory. The tests have not been submitted to the Food and Drug Administration (FDA) for clearance or approval and; therefore, are not FDA-licensed tests. The Molecular Analysis Laboratory is certified under the Clinical Laboratory Improvement Amendment (CLIA) of 1988 as qualified to perform high complexity clinical testing. The New York Blood Center has been approved by the New York State Department of Health to perform these tests under its current Clinical Laboratory Permit. These results are intended to predict a blood group antigen profile in a patient or donor, and are not intended for clinical diagnosis or as the sole means for patient management decisions. There are situations where testing DNA of a person may not reflect the red cell phenotype. Nucleotide changes that inactivate gene expression or rare new variant alleles may not be identified in these assays. Test results obtained from DNA isolated from leukocytes and other hematopoietic cells may differ from DNA isolated from other tissues in persons with a history of transplantation. 12 What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only

13 Cost of reagents for serological testing: Average from several centers Anti-C 1.15 Anti-E 1.15 Anti-c 1.15 Anti-e 2.20 Anti-K 1.55 Anti-k Anti-Kp a Anti-Kp b Anti-Fy a 7.11 Anti-Fy b Anti-Jk a Anti-Jk b Anti-S Anti-s 8.63 Anti-M 3.56 Anti-N 3.74 Anti-Di a Total $ Supplies, positive and negative controls, and labor significantly increases this cost Total cost: >$150-$ What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only

14 Goals of DNA-based testing Minimize (eliminate) alloimmunization Provide more precise matched blood versus least incompatible when patients have warm autoantibodies Reduce cost: Decrease patient work-ups: - EGA treatment; hypotonic wash - Adsorption/elution Locating rare blood These costs have not been previously calculated but offer significant savings. 14 What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only

15 In the year More personalized health care Wikipedia definition: Personalized medicine is a medical model that proposes the customization of healthcare, with decisions and practices being tailored to the individual patient by use of genetic or other information* Genotype all patients Only needs to be done once for a patient Results are part of the patient s electronic medical record *Personalized Medicine. (2009, March 16). In Wikipedia, The Free Encyclopedia. Retrieved September 28, 2012, from 15 What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only

16 Next generation sequencing Sequencing of the whole genome 23 and me Submit saliva and get results on numerous SNPs for only $299 Connect to ancestry Take a more active role in managing your health Third generation sequencing Predicted to sequence whole genome for a few hundred dollars in matter of hours. Analysis of all the sequence results is still ongoing 16 What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only

17 QUESTIONS 17 What s Next? Sunitha Vege 8 Oct 2012 Next-Generation Red Cell Typing Business Use Only