Related Experiment Video
Updated: Jun 10, 2026
Methods to Increase the Sensitivity of High Resolution Melting Single Nucleotide Polymorphism Genotyping in Malaria
Published on: November 10, 2015
Improved genotyping of the human minor histocompatibility antigen HB-1 by polymerase chain reaction with
1Department of Microbiology, College of Medicine, The Catholic University of Korea, Seocho-Gu, Seoul, Korea.
Abstract:
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.
