title: High-resolution melting analysis as an alternative method for human neutrophil antigen genotyping.
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.
authors: PhD (corresponding author), Deputy Director of Preparation Development.
authors: Kazuta
authors: K
authors: Yasui
authors: PhD, Deputy Director of Laboratory Development.
authors: Mitsunobu
authors: M
authors: Tanaka
authors: PhD.
authors: Tomoya
authors: T
authors: Hayashi
authors: MT, Deputy Director of 3rd Laboratory.
authors: Nobuki
authors: N
authors: Matsuyama
authors: PhD.
authors: Ayumu
authors: A
authors: Kuroishi
authors: PhD, Director of Preparation Development.
authors: Rika A
authors: RA
authors: Furuta
authors: MD, PhD, Deputy Director General.
authors: Yoshihiko
authors: Y
authors: Tani
authors: MD, PhD, Senior Director of Quality Control, Japanese Red Cross Kinki Block Blood Center, 7-5-17, Asagi Saito, Ibaraki, Osaka 567-0085 Japan.
authors: Fumiya
authors: F
authors: Hirayama
mesh_terms: D044466:Asian People; D005260:Female; D005787:Gene Frequency; D060005:Genotyping Techniques; D006801:Humans; D007519:Isoantigens / Q000235:genetics*; D007564:Japan; D008297:Male; D009504:Neutrophils; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide
chemical_list: D007519:Isoantigens