Microarray Beads for Identifying Blood Group Single Nucleotide Polymorphisms

• Published in: Transfusion medicine and hemotherapy Vol. 36; no. 3; pp. 157 - 160
• Main Authors: Drago, Francesca, Karpasitou, Katerina, Poli, Francesca
• Format: Journal Article
• Language: English
• Published: Basel, Switzerland S. Karger GmbH 01.01.2009
• Subjects: Review Article · Übersichtsarbeit; Review · Übersichtsarbeit; SNPs; Blood group; Single nucleotide polymorphism; Micrarray beads
• ISSN: 1660-3796; 1660-3818
• DOI: 10.1159/000215707

We have developed a high-throughput system for single nucleotide polymorphism (SNP) genotyping of alleles of diverse blood group systems exploiting Luminex technology. The method uses specific oligonucleotide probes coupled to a specific array of fluorescent microspheres and is designed for typing Jk a /Jk b , Fy a /Fy b , S/s, K/k, Kp a /Kp b , Js a /Js b , Co a /Co b and Lu a /Lu b alleles. Briefly, two multiplex PCR reactions (PCR I and PCR II) according to the laboratory specific needs are set up. PCR I amplifies the alleles tested routinely, namely Jk a /Jk b , Fy a /Fy b , S/s, and K/k. PCR II amplifies those alleles that are typed less frequently. Biotinylated PCR products are hybridized in a single multiplex assay with the corresponding probe mixture. After incubation with R-phycoerythrinconjugated streptavidin, the emitted fluorescence is analyzed with Luminex 100. So far, we have typed more than 2,000 subjects, 493 of whom with multiplex assay, and there have been no discrepancies with the serology results other than null and/or weak phenotypes. The cost of consumables and reagents for typing a single biallelic pair per sample is less than EUR 3.–, not including DNA extraction costs. The capability to perform multiplexed reactions makes the method markedly suitable for mass screening of red blood cell alleles. This genotyping approach represents an important tool in transfusion medicine.