Development and Validation of a Fully Automated Platform for Extended Blood Group Genotyping

• Published in: The Journal of molecular diagnostics : JMD Vol. 18; no. 1; pp. 144 - 152
• Main Authors: Boccoz, Stephanie A, Le Goff, Gaelle C, Mandon, Celine A, Corgier, Benjamin P, Blum, Loïc J, Marquette, Christophe A
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 2016; American Society for Investigative Pathology (ASIP)
• Subjects: Biochemistry; Biochemistry, Molecular Biology; Blood Group Antigens - genetics; Blood Group Antigens - immunology; Blood Grouping and Crossmatching - methods; Chemical Sciences; Erythrocytes - immunology; Genotyping Techniques - methods; Humans; Life Sciences; Multiplex Polymerase Chain Reaction; Oligonucleotide Array Sequence Analysis; Organic chemistry; Pathology; Polymorphism, Single Nucleotide - genetics
• ISSN: 1525-1578; 1943-7811
• DOI: 10.1016/j.jmoldx.2015.09.002

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.