Molecular basis for D− Japanese: identification of novel DEL and D− alleles. Issue 4 (7th May 2015)

• Record Type: Journal Article
• Authors: Ogasawara, K.; Suzuki, Y.; Sasaki, K.; Osabe, T.; Isa, K.; Tsuneyama, H.; Uchikawa, M.; Satake, M.; Tadokoro, K.
• Is Part Of: Vox sanguinis. Volume 109:Issue 4(2015)
• Page Start: 359
• Page End: 365
• Publication Date: 2015-05-07
• Subjects: Blood -- Periodicals; Blood -- Transfusion -- Periodicals; Immunohematology -- Periodicals; Immunopathology -- Periodicals; 615.39
• DOI: 10.1111/vox.12290
• Languages: English
• ISSNs: 0042-9007

Abstract

<abstract abstract-type="main" id="vox12290-abs-0001"> <title> <x xml:space="preserve">Abstract</x> </title> <sec id="vox12290-sec-0001" sec-type="section"> <title>Background and Objectives</title> <p>The occurrence of D− is approximately 0·5% in Japanese, but DEL in apparently D− individuals is relatively common compared with that in Caucasian populations. On the basis of molecular genetics, we examined D− Japanese blood donors.</p> </sec> <sec id="vox12290-sec-0002" sec-type="section"> <title>Methods</title> <p>A standard serological technique was used for RhD typing, and we selected 3526 D− blood samples. Genomic DNA obtained from whole blood was used for <italic>RHD</italic> analysis by polymerase chain reaction (PCR) and sequencing. Multiplex PCR to detect all of the <italic>RHD</italic> exons and use of PCR‐sequence‐specific primer (PCR‐SSP) to detect <italic>RHD</italic> deletion (<italic>RHD*01N.01</italic>) and c.1227G>A mutation (for <italic>RHD*01EL.01</italic>) were performed.</p> </sec> <sec id="vox12290-sec-0003" sec-type="section"> <title>Results</title> <p>Multiplex PCR and PCR‐SSP revealed that 3091 of 3526 D− individuals (87·7%) were homozygous for <italic>RHD*01N.01</italic>, and 318 individuals (9·0%) had the <italic>RHD*01EL.01/RHD*01N.01</italic> or <italic>RHD*01EL.01/RHD*01EL.01</italic> genotype. The other 103 in the 3526 individuals (2·9%) had the known <italic>D‐CE‐D</italic> hybrid allele, <italic>RHD*01N.04</italic>, and the association of <italic>RHCE*Ce</italic> with <italic>RHD*01EL.01</italic> as well as <italic>RHD*01N.04</italic> was observed. The remaining 14 individuals had <italic>RHD*01N.01</italic> hemizygous with one of the following alleles: <italic>RHD*01N.06</italic> (3), <italic>RHD*01N.07</italic> (1), <italic>RHD*04N.01</italic> (1), <italic>RHD*DEL8</italic> (1), <italic>RHD</italic> with c.761C>G (p.Ser254Ter) (2), <italic>RHD</italic> with c.1252T>A (p.Ter418Lysex26) (2) and apparently common <italic>RHD</italic> (4). Adsorption and elution tests with anti‐D revealed that the individuals with c.761C>G mutation were D− while the individuals with c.1252T>A mutation were DEL.</p> </sec> <sec id="vox12290-sec-0004" sec-type="section"> <title>Conclusions</title> <p>The <italic>RHD</italic> genotype of more than 96% of D− Japanese could be determined by conventional PCR‐SSP. In addition, we identified a novel <italic>DEL</italic> allele having c.1252T>A mutation and a novel <italic>RHD</italic> silencing allele having c.761C>G nonsense mutation.</p> </sec> </abstract>