Abstract
Neutrophil antigens are implicated in a variety of clinical conditions, including neonatal immune neutropenia, transfusion-related acute lung injury, refractoriness to granulocyte transfusions, febrile transfusion reactions, and autoimmune neutropenia. In this report, we describe simultaneous genotyping of human neutrophil antigens (HNA)-1, -3, -4, and -5 using PCR with allele-specific TaqMan probes and end-point fluorescence detection, which is a robust, rapid, and reproducible method, allowing for high-throughput genotyping.
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References
Bux J, Jung KD, Kauth T et al (1992) Serological and clinical aspects of granulocyte antibodies leading to alloimmune neonatal neutropenia. Transfus Med 2:143–149
Bux J, Behrens G, Jäger G, Welte K (1997) Diagnosis and clinical course of autoimmune neutropenia in infancy: analysis of 240 cases. Blood 89:1027–1036
Bux J (2011) Antibody-mediated (immune) transfusion-related acute lung injury. Vox Sang 100:122–128
Ory PA, Clark MR, Kwoh EE, Clarkson SB, Goldstein IM (1989) Sequences of complementary DNAs that encode the NA1 and NA2 forms of Fc receptor III on human neutrophils. J Clin Invest 84:1688–1691
Bux J, Stein EL, Bierling P, Fromont P, Clay M, Stroncek D, Santoso S (1997) Characterization of a new alloantigen (SH) on the human neutrophil Fc gamma receptor IIIb. Blood 89:1027–1034
Ravetch JV, Perussia B (1989) Alternative membrane forms of Fc gamma RIII(CD16) on human natural killer cells and neutrophils. Cell type-specific expression of two genes that differ in single nucleotide substitutions. J Exp Med 170:481–497
Bux J (2008) Human neutrophil alloantigens. Vox Sang 94:277–285
Huizinga TWJ, Kleijer M, Tetteroo PAT, Roos D, von dem Borne AEGK (1990) Biallelic neutrophil NAantigen system is associated with a polymorphism on the phospho-inositol-linked Fcγ receptor III (CD16). Blood 75:213–217
Kutyavin IV, Afonina IA, Mills A et al (2000) 3′-minor groove binder-DNA probes increase sequence specificity at PCR extension temperatures. Nucleic Acids Res 28:655–661
Nielsen KR, Koelbaek MD, Varming K, Baech J, Steffensen R (2012) 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. Tissue Antigens 80:249–253
Flesch BK, Reil A, Bux J (2011) Genetic variation of the HNA-3a encoding gene. Transfusion 11:2391–2397
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© 2015 Springer Science+Business Media New York
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Steffensen, R., Baech, J., Nielsen, K.R. (2015). Allelic Discrimination by TaqMan-PCR for Genotyping of Human Neutrophil Antigens. In: Bugert, P. (eds) Molecular Typing of Blood Cell Antigens. Methods in Molecular Biology, vol 1310. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2690-9_17
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DOI: https://doi.org/10.1007/978-1-4939-2690-9_17
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2689-3
Online ISBN: 978-1-4939-2690-9
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