Determination of DRB alleles using PCR amplification and allele-specific primers.
HLA class-II allelic diversity is commonly defined using polymerase chain reaction (PCR) in combination with sequence-specific oligotyping (PCR-SSO) or the combination of PCR and restriction fragment length polymorphism methods (PCR-RFLP). Nevertheless, the identification of the DRB polymorphism by PCR-SSO is a time-consuming procedure and the PCR-RFLP is cumbersome. A rapid technique which allows a precise and extensive HLA-DRB typing is required, particularly in order to study the role of class-II matching in organ transplantation. A DRB typing method based on the detection and length of PCR products amplified using combination of allele specific primers has been developed. Thirty-four DRB alleles (29 DRB1, 4DRB3, 1DRB4) can be detected using 29 primers distributed into 19 amplification mixtures.
Lepage
V
V
Laboratoire d'Immunologie et Histocompatibilité, Hôpital St Louis, Paris.
Schaeffer
V
V
Mallet
C
C
Ivanova
R
R
Khalil
I
I
Charron
D
D
D000483:Alleles; D002460:Cell Line; D017931:DNA Primers; D006684:HLA-DR Antigens / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D017931:DNA Primers; D006684:HLA-DR Antigens