title: A method of HLA class II typing using nonradioactive labelled oligonucleotides.
abstract: The typing of HLA class II genes using molecular biology techniques has brought undoubtedly new insights in the analysis of their polymorphism. Particularly interesting is the dot-blot analysis of enzymatically-amplified genomic DNA hybridized with sequence-specific oligonucleotides. In order to use this technique of typing on a routine basis, we established a non-radioactive detection method of enzymatically-amplified genomic DNA dot-blots. We could clearly demonstrate that, using biotin-labelled specific oligonucleotides, it was possible to specifically discriminate between DQB1 first domain DNA sequences displaying three, two or even only one base-pair difference at a given codon position. The very satisfactory sensitivity level reached by this non-radioactive detection method could safely allow its use for clinical applications of HLA typing at the DNA level.
authors.affiliation: Laboratoire d'Immunologie-INSERM U 291, Hopital Saint-Eloi, Montpellier, France.
authors.forename: J F
authors.initials: JF
authors.lastname: Eliaou
authors.forename: M
authors.initials: M
authors.lastname: Humbert
authors.forename: P
authors.initials: P
authors.lastname: Balaguer
authors.forename: L
authors.initials: L
authors.lastname: Gebuhrer
authors.forename: S
authors.initials: S
authors.lastname: Amsellem
authors.forename: H
authors.initials: H
authors.lastname: Bétuel
authors.forename: J C
authors.initials: JC
authors.lastname: Nicolas
authors.forename: J
authors.initials: J
authors.lastname: Clot
mesh_terms: D004247:DNA / Q000235:genetics; D005784:Gene Amplification; D005819:Genetic Markers / Q000032:analysis; D006683:HLA-DQ Antigens / Q000235:genetics; D006579:Heterozygote; D000949:Histocompatibility Antigens Class II / Q000235:genetics*; D006720:Homozygote; D006801:Humans; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000032:analysis*; D012689:Sequence Homology, Nucleic Acid
chemical_list: D005819:Genetic Markers; D006683:HLA-DQ Antigens; D000949:Histocompatibility Antigens Class II; D009841:Oligonucleotides; D004247:DNA