The hybridization products obtained by PCR using sequence-specific oligonucleotides can be traced either by colorimetric (streptavidin–biotin)-, X-ray (digoxigenin–CSPD)-, or fluorescence (FITC, PE)-based detection systems. To achieve a faster, reliable, automated typing technique microbead and fluorescence detection technology have been combined and introduced to this field (XMAP™ technology). For each locus, a series of microspheres, which are recognizable by their specific color originating from two internal fluorescent dyes, are used. Each microsphere is coupled with a single probe that is capable of hybridizing with the biotin-labeled complementary amplicon. Once hybridization occurs, it can be quantified by measuring the fluorescence signal originating from fluorescently (streptavidin–PE) labeled amplicons captured by the beads. Currently, there are two commercially available systems that differ in the scale of probes and the methods used for amplification and denaturation. One of these is described in detail in this chapter.
PCR-SSO Luminex™ XMAP™ technology HLA typing
This is a preview of subscription content, log in to check access.
Springer Nature is developing a new tool to find and evaluate Protocols. Learn more
Blasczyk R (1998) New HLA typing methods. In: Huhn D (ed) New diagnostic methods in oncology and hematology. Springer, Berlin, pp 143–195CrossRefGoogle Scholar
Saiki RK, Bugawan TL, Horn GT, Mullis KB, Erlich HA (1986) Analysis of enzymatically amplified beta globin and HLA-DQ alpha DNA with allele specific oligonucleotide probes. Nature 324:163–166PubMedCrossRefGoogle Scholar
Kostyu DD, Pfohl J, Ward FE, Lee J, Murray A, Amos DB (1993) Rapid HLA-DR oligotyping by an enzyme-linked immunosorbent assay performed in microtiter trays. Hum Immunol 38:148–158PubMedCrossRefGoogle Scholar
Olerup O, Zetterquist H (1992) HLA-DRB1*01 subtyping by allele specific pcr amplification: a sensitive, specific and rapid technique. Tissue Antigens 37:197–204CrossRefGoogle Scholar
Bugawan TL, Apple R, Erlich HA (1994) A method for typing polymorphism at the HLA-A Locus using PCR amplification and immobilized oligonucleotide probes. Tissue Antigens 44:137–147PubMedCrossRefGoogle Scholar
Middleton D, Williams F, Cullen C, Mallon E (1995) Modification of an HLA-B PCR-SSOP typing system leading to improved allele determination. Tissue Antigens 45:232–236PubMedCrossRefGoogle Scholar
Kennedy LJ, Poulton KV, Dyer PA, Ollier WE, Thomson W (1995) Definition of HLA-C alleles using sequence specific oligonucleotide probes (PCR-SSOP). Tissue Antigens 46: 187–195PubMedCrossRefGoogle Scholar
Cereb N, Maye P, Lee S, Kong Y, Yang SY (1995) Locus specific amplification of HLA Class I genes from genomic DNA: locus specific sequences in the first and third introns of HLA-A, -B and -C, alleles. Tissue Antigens 45: 1–11PubMedCrossRefGoogle Scholar
Robinson J, Malik, Parham P, Bodmer JG, Marsh SGE (2000) IMGT/HLA database a sequence database for the human major histocompatibility complex. Tissue Antigens 55: 280–287PubMedCrossRefGoogle Scholar
Wu YY, Csako G (2006) Rapid and/or high throughput genotyping for human red blood cell, platelet, and leukocyte antigens, and forensic applications. Clin Chim Acta 363: 165–176PubMedCrossRefGoogle Scholar
Dunbar AS (2006) Applications of Luminex® MAP technology for rapid, high throughput multiplexed nucleic acid detection. Clin Chim Acta 363:71–82PubMedCrossRefGoogle Scholar
Fulton RJ, McDade RL, Smith PL, Kienker LJ, Kettman JR Jr (1997) Advanced multiplexed analysis with FlowMetrix system. Clin Chem 43:1749–1756PubMedGoogle Scholar