Improved genotyping of the human minor histocompatibility antigen HB-1 by polymerase chain reaction with sequence-specific primers using a complementary oligonucleotide.
Single nucleotide polymorphisms of minor histocompatibility antigens (mHags) have been genotyped by allele-specific polymerase chain reaction with sequence-specific primers (PCR-SSP). Because discriminating the genotype of HB-1 Y by PCR-SSP under various PCR conditions was difficult, we optimized the use of oligonucleotides complementary to the allele-specific forward primer to improve the specificity of the HB-1 Y PCR-SSP. Specific allele discrimination was possible with an annealing temperature between 61°C and 63°C and in the presence of a threefold excess of a 15-bp complementary oligonucleotide. In conclusion, the inclusion of a complementary oligonucleotide in the PCR-SSP assay may improve its specificity and selectivity for genotyping several mHags for which optimizing PCR conditions have been difficult.
Park
M-J
MJ
Department of Microbiology, College of Medicine, The Catholic University of Korea, Seocho-Gu, Seoul, Korea.
Choi
H-B
HB
Kim
T-G
TG
D000483:Alleles; D017931:DNA Primers / Q000235:genetics* / Q000276:immunology; D005838:Genotype; D015235:HLA-B Antigens / Q000235:genetics* / Q000276:immunology; D006801:Humans; D015778:Minor Histocompatibility Antigens; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D015235:HLA-B Antigens; C117270:HMHB1 protein, human; D015778:Minor Histocompatibility Antigens