title: Development and Validation of a Fully Automated Platform for Extended Blood Group Genotyping.
abstract: Thirty-five blood group systems, containing >300 antigens, are listed by the International Society of Blood Transfusion. Most of these antigens result from a single nucleotide polymorphism. Blood group typing is conventionally performed by serology. However, this technique has some limitations and cannot respond to the growing demand of blood products typed for a large number of antigens. The knowledge of the molecular basis of these red blood cell systems allowed the implementation of molecular biology methods in immunohematology laboratories. Here, we describe a blood group genotyping assay based on the use of TKL immobilization support and microarray-based HIFI technology that takes approximately 4 hours and 30 minutes from whole-blood samples to results analysis. Targets amplified by multiplex PCR were hybridized on the chip, and a revelation step allowed the simultaneous identification of up to 24 blood group antigens, leading to the determination of extended genotypes. Two panels of multiplex PCR were developed: Panel 1 (KEL1/2, KEL3/4; JK1/2; FY1/2; MNS1/2, MNS3/4, FY*Fy et FY*X) and Panel 2 (YT1/2; CO1/2; DO1/2, HY+, Jo(a+); LU1/2; DI1/2). We present the results of the evaluation of our platform on a panel of 583 and 190 blood donor samples for Panel 1 and 2, respectively. Good correlations (99% to 100%) with reference were obtained.
authors.affiliation: Institute of Chemistry and Molecular and Supramolecular Biochemistry Team Enzyme Engineering, Biomimetic Membranes and Supramolecular Assemblies, CNRS 5246 ICBMS, Université Lyon 1, Villeurbanne, France.
authors.forename: Stephanie A
authors.initials: SA
authors.lastname: Boccoz
authors.affiliation: AXO Science SAS, Villeurbanne, France.
authors.forename: Gaelle C
authors.initials: GC
authors.lastname: Le Goff
authors.affiliation: AXO Science SAS, Villeurbanne, France.
authors.forename: Celine A
authors.initials: CA
authors.lastname: Mandon
authors.affiliation: AXO Science SAS, Villeurbanne, France.
authors.forename: Benjamin P
authors.initials: BP
authors.lastname: Corgier
authors.affiliation: Institute of Chemistry and Molecular and Supramolecular Biochemistry Team Enzyme Engineering, Biomimetic Membranes and Supramolecular Assemblies, CNRS 5246 ICBMS, Université Lyon 1, Villeurbanne, France.
authors.forename: Loïc J
authors.initials: LJ
authors.lastname: Blum
authors.affiliation: Institute of Chemistry and Molecular and Supramolecular Biochemistry Team Enzyme Engineering, Biomimetic Membranes and Supramolecular Assemblies, CNRS 5246 ICBMS, Université Lyon 1, Villeurbanne, France. Electronic address: christophe.marquette@univ-lyon1.fr.
authors.forename: Christophe A
authors.initials: CA
authors.lastname: Marquette
mesh_terms: D001789:Blood Group Antigens / Q000235:genetics* / Q000276:immunology; D001788:Blood Grouping and Crossmatching / Q000379:methods*; D004912:Erythrocytes / Q000276:immunology; D060005:Genotyping Techniques / Q000379:methods*; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D020411:Oligonucleotide Array Sequence Analysis; D020641:Polymorphism, Single Nucleotide / Q000235:genetics
chemical_list: D001789:Blood Group Antigens