<abstract xmlns="http://www.w3.org/1999/xhtml"><p>Human neutrophil antigen (HNA)-typed granulocyte panels are widely used to screen for the presence of HNA antibodies and to determine antibody specificity. Many laboratories screen donors for HNA genotypes using low-throughput methods such as allele-specific polymerase chain reaction (PCR), PCR–restriction fragment–length polymorphism, and multiplex PCR. In the present study, we used a high-resolution melting (HRM) analysis to determine HNA genotypes. For the HRM analysis, purified genomic DNA samples were amplified via PCR with HNA-specific primers. Nucleotide substitutions in genes encoding HNAs were differentiated on the basis of the HRM curves, and the results of HRM and DNA sequencing analyses were determined to be in complete agreement. The gene frequency of HNA-1a, -1b, -1c, -3a, -3b, -4a, -4b, -5a, and -5b in the Japanese population was consistent with the previous reports. Our results suggest that HRM analysis can be used for genotyping HNA antigens determined by single nucleotide substitutions. <italic><bold>Immunohematology</bold></italic><bold>2015;31:7–13.</bold></p></abstract>

Human neutrophil antigen (HNA)-typed granulocyte panels are widely used to screen for the presence of HNA antibodies and to determine antibody specificity. Many laboratories screen donors for HNA genotypes using low-throughput methods such as allele-specific polymerase chain reaction (PCR), PCR–restriction fragment–length polymorphism, and multiplex PCR. In the present study, we used a high-resolution melting (HRM) analysis to determine HNA genotypes. For the HRM analysis, purified genomic DNA samples were amplified via PCR with HNA-specific primers. Nucleotide substitutions in genes encoding HNAs were differentiated on the basis of the HRM curves, and the results of HRM and DNA sequencing analyses were determined to be in complete agreement. The gene frequency of HNA-1a, -1b, -1c, -3a, -3b, -4a, -4b, -5a, and -5b in the Japanese population was consistent with the previous reports. Our results suggest that HRM analysis can be used for genotyping HNA antigens determined by single nucleotide substitutions. Immunohematology2015;31:7–13.

High-resolution melting analysis as an alternative method for human neutrophil antigen genotyping

Immunohematology

Human neutrophil antigen (HNA)-typed granulocyte panels are widely used to screen for the presence of HNA antibodies and to determine antibody specificity....

High-resolution melting analysis as an alternative method for human neutrophil antigen genotyping

Article Category: Original Report

Publié en ligne: 26 oct. 2019

Pages: 7 - 13

DOI: https://doi.org/10.21307/immunohematology-2019-064

Mots clés
neutrophil, HNA, antibody, genotyping, high-resolution melting analysis

© 2015 K. Yasui et al., published by Sciendo

High-resolution melting analysis as an alternative method for human neutrophil antigen genotyping

Article Category: Original Report

Publié en ligne: 26 oct. 2019

Pages: 7 - 13

DOI: https://doi.org/10.21307/immunohematology-2019-064

Mots clésneutrophil, HNA, antibody, genotyping, high-resolution melting analysis

© 2015 K. Yasui et al., published by Sciendo

Mots clés
neutrophil, HNA, antibody, genotyping, high-resolution melting analysis