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Tissue Antigens. 2010 Dec;76(6):482-6. doi: 10.1111/j.1399-0039.2010.01546.x. Epub 2010 Aug 16.

Improved genotyping of the human minor histocompatibility antigen HB-1 by polymerase chain reaction with sequence-specific primers using a complementary oligonucleotide.

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Department of Microbiology, College of Medicine, The Catholic University of Korea, Seocho-Gu, Seoul, Korea.


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.

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