A fast and easy real-time PCR genotyping method for the HLA-G 14-bp insertion/deletion polymorphism in the 3' untranslated region.
Human leukocyte antigen-G (HLA-G) is a non-classical HLA class Ib molecule shown to exhibit immunomodulatory function in a wide range of immune-based disorders. A number of functional HLA-G gene polymorphisms have been identified, including a 14-bp insertion/deletion polymorphism in exon 8 of the 3' untranslated region of the HLA-G gene, which has been associated with HLA-G mRNA stability. Moreover, studies show that homozygosity for the 14-bp insertion/deletion polymorphism is associated with lower HLA-G mRNA and protein levels and unique alternative splicing patterns. Here, we introduce a quick and reliable method to screen for the HLA-G 14-bp insertion/deletion polymorphism using an optimized real-time polymerase chain reaction protocol. The genotyping assay has been validated by comparison with conventional methods. As results can be obtained within a few hours, the assay will have a potential for clinical use.
Department of Clinical Biochemistry, Centre for Immune Regulation and Reproductive Immunology, Copenhagen University Hospital, DK-4000 Roskilde, Denmark.
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D020413:3' Untranslated Regions / Q000235:genetics*; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D060005:Genotyping Techniques / Q000379:methods*; D059951:HLA-G Antigens / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D011110:Polymorphism, Genetic; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D017384:Sequence Deletion; D013997:Time Factors
D020413:3' Untranslated Regions; D059951:HLA-G Antigens