SBIR-STTR Award

Low-cost nucleic acid analysis, to detect HIV &HBV, has revolutionized screening of the blood supply. In spite of that new genetic sophistication, traditional blood group typing [ABO, Rh] is still performed, for the most part, via variations upon 80-year-old agglutination testing. Recently, the genetic variation which gives rise to the standard blood groups has been defined, and shown to be relatively simple. In parallel, it has been shown that, to enhance transfusion quality, the historical marker set might be expanded to include a more complete panel of 7 traits: [ABO, Rh, Duffy, Kidd, Kell, Dombrock &MNS] for which the underlying genetics are now known. These two parallel trends suggest that genetic testing has emerged as the future of blood group screening: but only if the genetic test can be delivered at roughly the same cost and level of technical simplicity as agglutination, or the current panel of nucleic acid based pathogen tests. We have observed that, with minor modification, the suite of biochemical, hardware &software technologies that we have developed for microarray based HLA-typing on raw blood could, instead, be transformed into low-cost technologies for blood-group typing, which we refer to as ""The Transfusion-Chip"". In this 6-month Phase I plan, we propose 2 Specific Aims as Milestones to justify a subsequent Phase II. SA1. Design &preliminary validation of 13-15 PCR reactions to be performed in parallel. All informative sites among the set of 7 blood marker genes will be amplified in parallel, to generate a dye-labeled amplicon set that is ready to be used for microarray hybridization. SA2. Microarray probe design, fabrication &preliminary validation. A Transfusion-Chip prototype will be designed and fabricated, to interrogate the informative sequence variations which define blood group variation among the set of 7 loci. The validation will employ amplified DNAs from SA1. Relation to a Follow-on Phase II. Completion of Phase I will yield a prototype PCR-based sample labeling reaction (that works on raw blood) and a prototype low-cost Transfusion-Chip microarray for DNA based blood-typing. Phase II will focus on refinement of the PCR and microarray designs, manufacturing scale-up, preliminary PCR kit fabrication and delivery of those materials to beta testers, comprising 3-4 top blood labs: to compare Transfusion-Chip performance to serological typing and to the competing microarray and PCR tests. Public Health Narrative. Genetic testing for pathogen contamination has revolutionized the safety of the blood supply, yet analogous DNA based testing of the underlying blood groups has lagged-behind, due to cost and complexity. A microarray-based approach to blood group typing that we propose here, will allow blood type analysis to be modernized, by exploiting two decades worth of blood group genetics: allowing analysis of blood group antigen variation, in a way that is more complete than can be obtained by serology, at a cost and level of technical simplicity that is as practical as that of blood pathogen screening. The resulting technology will not only reduce the cost of blood supply testing in the US, but we propose, will enable a new generation of enhanced, DNA-based blood group typing in the developing world.

Public Health Relevance:
A microarray-based approach to blood group typing that we propose here, will allow blood type analysis to be modernized, by exploiting two decades worth of blood group genetics: allowing analysis of blood group antigen variation, in a way that is more complete than can be obtained by serology, at a cost and level of technical simplicity that is as practical as that of blood pathogen screening. The resulting technology will not only reduce the cost of blood supply testing in the US, but we propose, will enable a new generation of enhanced, DNA-based blood group typing in the developing world.

Thesaurus Terms:
Aids Virus;Acquired Immune Deficiency Syndrome Virus;Acquired Immunodeficiency Syndrome Virus;Adopted;Agglutination;Agglutination Tests;Antigen Variation;Antigenic Variability;Antigenic Variation;Biochemical;Blood;Blood Group Antigens;Blood Grouping;Blood Reticuloendothelial System;Blood Sample;Blood Typing;Blood Groups;Blood Specimen;Blood Typing Procedure;Bypass;Coloring Agents;Computer Software;Dna;Dna Chips;Dna Microarray;Dna Microarray Chip;Dna Microchips;Dna Purification;Deoxyribonucleic Acid;Dyes;Ensure;Future;Gene Variant;Generations;Genes;Genetic;Genetic Diversity;Genetic Variation;Genetic Screening Method;Hbv;Hiv;Htlv-Iii;Hepatitis B Virus;Homologous Serum Hepatitis Virus;Human Immunodeficiency Viruses;Human T-Cell Leukemia Virus Type Iii;Human T-Cell Lymphotropic Virus Type Iii;Human T-Lymphotropic Virus Type Iii;Lav-Htlv-Iii;Label;Lymphadenopathy-Associated Virus;Manuals;Measures;Microarray Analysis;Microarray-Based Analysis;Minor;Modification;Niaid;National Institute Of Allergy And Infectious Disease;Nucleic Acids;Performance;Phase;Population;Process;Public Health;Reaction;Reagent;Robotics;Sbir;Sbirs (R43/44);Safety;Sampling;Screening Procedure;Serologic;Serologic Tests;Serological;Serological Tests;Site;Small Business Innovation Research;Small Business Innovation Research Grant;Software;Solutions;Sorting - Cell Movement;Trnsf;Technology;Testing;Transfusion;Validation;Variant;Variation;Variation (Genetics);Vascular Blood Supply;Viral;Virus-Hiv;Work;Agglutination Reaction;Allelic Variant;Base;Blood Supply;Computer Program/Software;Cost;Design;Designing;Genetic Technology;Genetic Testing;Manufacturing Scale-Up;Microarray Technology;Pathogen;Prototype;Public Health Medicine (Field);Scale Up;Screening;Screenings;Serology;Sorting;Trait;Trend;Vascular Supply

Blood-Typing is the progenitor of the entire field of personalized medicine. In the recently awarded 6-month Phase I, we proposed to take the first step in the development of an extremely-low-cost microarray test, "The Transfusion-Chip" (T-Chip) which convert the historical practice of Blood-Typing by serology into a simple inexpensive DNA test. Here, in this follow-on Phase II, we demonstrate substantial success in achieving key Phase I milestones: we have developed and optimized 22 PCR reactions, comprising the entire 7 gene blood group gene set [ABO, Rh, Duffy, Kidd, Kell, Dombrock & MNS]; we have fabricated a Transfusion-Chip microarray prototype capable of analyzing all known genetic variation in the 7 gene set; and via technology that we had not yet invented when we filed the Phase I, we have developed a new variant of the original GenUSA microarray test which will drop the manufacturing cost of T-Chips to values as low as $1 per microarray. In Phase I we have also shown that the raw, unpurified OrageneTM-stabilized saliva can used as the basis for DNA based Blood-Typing on the T-Chip. We feel that the coupling of raw OrageneTM-stabilized saliva collection with the low cost T-Chip could become a unique technology pairing: because it would allow self-collection of DNA samples, at home, then low- cost ambient temperature shipping to centralized labs, to be followed by very-low-cost, very high throughput Blood-Typing on T-Chips. This model for population scale Blood-Typing will be the development focus in this Phase II: thereby enabling, we propose, an entirely new vision for population-scale blood banking & transfusion medicine, and in the process, will allow Blood- Typing to return, in the 21st century, to prominence as the model and gold standard for the field of genetically personalized medicine: as it first did a hundred years ago. Based on that model for population-scale blood group genetic testing, we propose the following set of Phase II Aims. SA1. Complete & Validate the Multiplex PCR Front-end to the Transfusion-Chip, for DNA Samples. SA2. Complete & Validate the Multiplex PCR Front-end to the Transfusion-Chip, for Raw Samples. SA3. Optimize & validate fabrication of the Transfusion-Chip in the 48 array per slide "$1 microarray" format. SA4. Modify existing GenUSA analysis software, RicimerTM, for automated blood group typing. SA5. Validate the Transfusion-Chip for purified DNA, raw blood & raw OrageneTM-stabilized saliva samples. Phase II Deliverables & Commercialization Plan. Upon completion of this Phase II, we will have ready a Transfusion-Chip kit prototype (microarrays, PCR reagents, Hybridization Reagents, Specialized Software) for subsequent Beta Testing. This kit will be prepared under manufacturing control (ISO 13845) similar to the HLA-Chip products from GenUSA which are currently being launched into tissue compatibility and immunogenetics markets. Subsequent to completion of Phase II, GenUSA will launch the T-Chip into the RUO Blood-Typing market and prepare simultaneously for FDA 510(k) submission to enter into the diagnostics market either alone, or in partnership with those with a proven market presence in transfusion medicine or at-home sample collection, or both.

Public Health Relevance Statement:


Public Health Relevance:
Blood group antigen typing by serology (Blood-Typing) has been, for a century, the progenitor and gold standard for the entire field of personalized medicine. Here we propose to pair two technologies recently developed at GenUSA [Raw Sample Genotyping + $1 Microarrays] to allow Blood-Typing to become a low-cost genetic test: A saliva or a dried blood sample could be self-collected at home in a simple clinic, then shipped by ordinary book-rate mail to a central lab, where upon arrival in the lab, Blood-Typing would be performed inexpensively via "$1 microarray testing". We propose that this new vision for very-large-scale public health genetics will not only revolutionize the regulation of the blood supply and the practice of transfusion medicine, but will also serve as a model for a new approach to genetically personalized medicine: much as Blood-Typing, by serology, created (personalized) Transfusion Medicine a century ago.

Project Terms:
Antigenic Variation; Antigens; Award; base; Blood; Blood Banks; Blood Group Antigens; Blood groups; Blood specimen; Blood Transfusion; Blood typing procedure; Books; Buffers; Clinic; Collection; commercialization; Companions; Computer software; cost; Coupling; Databases; Development; Diagnostic; Diagnostic tests; DNA; DNA Microarray Chip; Drops; Funding; Genes; Genetic; Genetic screening method; Genotype; genotyping technology; Gold; Histocompatibility; Home environment; Immunogenetics; International; Mails; Marketing; Medicine; Microarray Analysis; Modeling; National Institute of Allergy and Infectious Disease; new technology; novel strategies; Phase; Population; population based; Process; progenitor; Program Development; programs; Progress Reports; prototype; public health medicine (field); public health relevance; Reaction; Reagent; Regulation; Rest; Saliva; sample collection; Sampling; Serologic tests; Shipping; Ships; Slide; Spottings; success; Technology; technology validation; Temperature; Testing; Transfusion; Variant; Variation (Genetics); Vascular blood supply; Vision