Improved KIR gene and HLA-C KIR ligand sequence-specific primer polymerase chain reaction genotyping using whole genome amplification.
Molecular analysis of genetic polymorphism for clinical or research purposes may be compromised by genomic DNA of limited quality and quantity. In this study, we have successfully tested the feasibility of using whole genome amplification (WGA) to allow genotyping for killer cell immunoglobulin-like receptor (KIR) genes and human leucocyte antigen (HLA)-C KIR ligand dimorphism on HLA-C. WGA was achieved by multiple displacement amplification (MDA) using bacteriophage phi29 polymerase. For KIR genotyping, a revised sequence-specific primer polymerase chain reaction protocol consisting of 23 primer pairs was used avoiding hitherto undetected cross-priming involving KIR2DL1, KIR2DS1, KIR3DL1 and KIR3DS1 alleles. Similarly, MDA-amplified genomic DNA was analyzed for the detection of the HLA-C KIR ligand groups C1 and C2, based on the amino acid K/N dimorphism in position 80.
Chainonthee
W
W
Medizinische Klinik u. Poliklinik I, Universitätsklinikum Carl Gustav Carus der Technischen Universität Dresden, Dresden, Germany.
Böttcher
G
G
Gagne
K
K
Füssel
M
M
Bignon
J-D
JD
Wassmuth
R
R
D000483:Alleles; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D015343:DNA Probes, HLA / Q000235:genetics; D055106:Genome-Wide Association Study; D005838:Genotype; D015236:HLA-C Antigens / Q000235:genetics*; D006801:Humans; D008024:Ligands; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D054340:Receptors, KIR / Q000235:genetics*; D054342:Receptors, KIR2DL1 / Q000235:genetics; D054341:Receptors, KIR3DL1 / Q000235:genetics; D054349:Receptors, KIR3DS1 / Q000235:genetics
D017931:DNA Primers; D015343:DNA Probes, HLA; D015236:HLA-C Antigens; C093100:KIR2DL1 protein, human; C496212:KIR2DS1 protein, human; C515083:KIR3DL1 protein, human; D008024:Ligands; D054340:Receptors, KIR; D054342:Receptors, KIR2DL1; D054341:Receptors, KIR3DL1; D054349:Receptors, KIR3DS1