A method of HLA class II typing using nonradioactive labelled oligonucleotides.
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.
Eliaou
J F
JF
Laboratoire d'Immunologie-INSERM U 291, Hopital Saint-Eloi, Montpellier, France.
Humbert
M
M
Balaguer
P
P
Gebuhrer
L
L
Amsellem
S
S
Bétuel
H
H
Nicolas
J C
JC
Clot
J
J
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
D005819:Genetic Markers; D006683:HLA-DQ Antigens; D000949:Histocompatibility Antigens Class II; D009841:Oligonucleotides; D004247:DNA