Tanaka M, Kamada I, Takahashi J, Hirayama F, Tani Y
Japanese Red Cross, Kinki Block Blood Center, Osaka, Japan.
Transfus Med. 2014 Feb;24(1):39-44. doi: 10.1111/tme.12085. Epub 2013 Oct 24.
Blood-group genotyping arrays have been widely used in Caucasian and African American populations, but have not been thoroughly tested in Japanese subjects.
To evaluate, using the BLOODchip(®) Reference genotyping system, the concordance of previously typed samples with expected phenotypes and the coverage of the Japanese variants.
Blood samples from 100 Japanese donors were obtained. DNA was extracted with QIAsymphony (Qiagen, Hilden, Germany). Samples were typed by serological methods and processed with the BLOODchip(®) . When a non-concordant result was identified, further sequencing by polymerase chain reaction-single specific primer (PCR-SSP) was performed.
Concordance between systems was 98% (736/751), and 98.8% (742/751) if only non-software-related non-concordances were considered. In the ABO group, 6 'No Call' (NC, inability of the BLOODchip(®) to assign a result) were ascribed to a variant of blood subtype A1 (A102; 467C>T), a common subtype in Asian populations, whereas three NC presented additional polymorphisms not contained in the BLOODchip(®) (A102/A205, A102/O06 and A204/O02). In the RhD group, one discrepancy was correctly genotyped as RHD1227A (Del phenotype) by the BLOODchip(®) (phenotyped as partial D, RHDDIVb). Another was phenotyped as D+ by the BLOODchip(®) (phenotyped weak D by serology) and confirmed as RHDD-CE(2)-D heterozygous by sequencing. The 3 RhD NC can be solved by further software update. For RhCE, one discrepancy was correctly genotyped for both systems; however, only the BLOODchip(®) was able to detect RHCECX allele.
By programming the A102 ABO variant into the system software with the new allele combinations, the BLOODchip(®) Reference is a suitable genotyping tool to be applied to Asian samples.
血型基因分型芯片已在白种人和非裔美国人中广泛应用，但尚未在日本人群中进行全面测试。
使用BLOODchip（®）参考基因分型系统，评估先前分型样本与预期表型的一致性以及日本变异的覆盖范围。
采集100名日本献血者的血液样本。用QIAsymphony（德国希尔德的Qiagen公司）提取DNA。样本通过血清学方法分型并用BLOODchip（®）处理。当发现不一致结果时，通过聚合酶链反应 - 单特异性引物（PCR - SSP）进行进一步测序。
两个系统之间的一致性为98%（736/751），如果仅考虑与软件无关的不一致情况，则为98.8%（742/751）。在ABO血型组中，6个“无结果（NC，BLOODchip（®）无法给出结果）”归因于血液亚型A1的一个变异（A102；467C>T），这是亚洲人群中的常见亚型，而3个NC表现出BLOODchip（®）中未包含的其他多态性（A102/A205、A102/O06和A204/O02）。在RhD血型组中，一个差异通过BLOODchip（®）被正确基因分型为RHD1227A（Del表型）（血清学表型为部分D，RHDDIVb）。另一个通过BLOODchip（®）表型为D+（血清学表型为弱D），经测序确认为RHDD - CE（2）-D杂合子。3个RhD无结果情况可通过进一步软件更新解决。对于RhCE，一个差异在两个系统中均被正确基因分型；然而，只有BLOODchip（®）能够检测到RHCECX等位基因。
通过将A102 ABO变异与新的等位基因组合编入系统软件，BLOODchip（®）参考系统是适用于亚洲样本的基因分型工具。
