title: Microfluidic approach to genotyping human platelet antigens.
abstract: Centralised laboratories routinely determine blood types by serological and molecular methods. Current practices have limitations in terms of cost, time and accessibility. Miniaturised microfluidic platforms offer an alternative to conventional genotyping methods, since they consume fewer reagents, provide faster analysis and allow for complete integration and automation. As these 'lab-on-a-chip' devices have been used for bacterial and viral detection, the authors investigated blood group genotyping as a novel application of microfluidic technology. To demonstrate the feasibility of microfluidic chip-based genotyping, the authors compared human platelet antigen 1 (HPA-1) genotype results from conventional and chip-based analysis for 19 blood donor specimens. DNA purification was performed with ChargeSwitch™ magnetic beads, DNA amplification (PCR), restriction length polymorphism (RFLP) and capillary electrophoresis (CE) for identification of the DNA on microfluidic chips. It was found that nine donors were HPA-1a/1a and ten were HPA-1a/1b. Concordance between the conventional and on-chip methods was achieved for all but one sample. All the steps were demonstrated for complete blood group genotyping analysis of patient whole blood specimens on separate microfluidic chips. Future work will focus on integration of all the genotyping protocols on a single microfluidic chip.
authors.affiliation: Canadian Blood Services, Research and Development, Edmonton, Canada.
authors.forename: C
authors.initials: C
authors.lastname: Lim
authors.forename: D P
authors.initials: DP
authors.lastname: Manage
authors.forename: A
authors.initials: A
authors.lastname: Atrazhev
authors.forename: G
authors.initials: G
authors.lastname: Denomme
authors.forename: C J
authors.initials: CJ
authors.lastname: Backhouse
authors.forename: J P
authors.initials: JP
authors.lastname: Acker
mesh_terms: D016824:Antigens, Human Platelet / Q000097:blood / Q000235:genetics*; D001483:Base Sequence; D004247:DNA / Q000097:blood* / Q000302:isolation & purification*; D017931:DNA Primers; D046209:Electrophoresis, Microchip; D060005:Genotyping Techniques; D006801:Humans; D039661:Integrin beta3; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity
chemical_list: D016824:Antigens, Human Platelet; D017931:DNA Primers; C061248:ITGB3 protein, human; D039661:Integrin beta3; C083451:human platelet antigen 1b; D004247:DNA