Frequencies of HNA-1, HNA-3, HNA-4, and HNA-5 in the Danish and Zambian populations determined using a novel TaqMan real time polymerase chain reaction method.
In this study, we report a novel real time polymerase chain reaction (Q-PCR) method using TaqMan probes for human neutrophil antigens (HNA)-1, -3, -4, and -5 genotyping. The method was validated in a Caucasian Danish population, a Zambian population, and in clinical samples using three different methods: an in-house polymerase chain reaction with sequence-specific primers (PCR-SSP) method, a commercial available PCR-SSP kit and a novel Q-PCR method. We observed no discrepancy in the genotype frequencies determined by the PCR-SSP methods and the TaqMan assay in the populations studied. In tests of a family of Nigerian origin and in samples carrying the rare SLC44A2*1:2 genotype, different results were produced by the commercial PCR-SSP kit and the real-time TaqMan assay. The TaqMan-based genotyping method was rapid and reproducible, allowing high-throughput HNA-1, -3, -4, and -5 genotyping.
Department of Clinical Immunology, Aalborg Hospital, Aalborg, Denmark. k.nielsen@rn.dk
K R
KR
Nielsen
M D
MD
Koelbaek
K
K
Varming
J
J
Baech
R
R
Steffensen
D003718:Denmark / Q000208:ethnology; D058851:GPI-Linked Proteins / Q000235:genetics; D005787:Gene Frequency / Q000235:genetics*; D005828:Genetics, Population; D060005:Genotyping Techniques; D006801:Humans; D007519:Isoantigens / Q000235:genetics* / Q000276:immunology; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D017452:Receptors, IgG / Q000235:genetics; D019914:Taq Polymerase / Q000378:metabolism*; D015024:Zambia / Q000208:ethnology
C496124:FCGR3A protein, human; C428200:FCGR3B protein, human; D058851:GPI-Linked Proteins; C518134:HNA-3a antigen, human; C531098:HNA-4A antigen; D007519:Isoantigens; D017452:Receptors, IgG; C067704:neutrophil-specific antigen NA1, human; D019914:Taq Polymerase