Tanaka M, Kamada I, Takahashi J, Kimura T, Tani Y
Japanese Red Cross Kinki Block Blood Center, Osaka, Japan.
Transfus Med. 2018 Jun;28(3):255-260. doi: 10.1111/tme.12474. Epub 2017 Sep 21.
The aim of this study was to evaluate the applicability of genotyping of the ABCG2 gene using MALDI-TOF MS and to estimate the allele frequency in the Japanese population.
Jr (a-) phenotype has a prevalence of approximately 0·05% among Japanese blood donors; DNA-based genotyping was conducted to investigate the molecular basis of the Jr (a-) phenotype along with serological typing. To detect all SNPs of the ABCG2 gene, a high-throughput SNP genotyping platform is needed.
Overall, 1004 Jr (a-) blood samples were collected from blood donors in Japan and pre-genotyped. To detect the SNPs of the ABCG2 gene using MALDI-TOF MS, polymerase chain reaction and unextend primer were designed. In total, 205 Jr (a-) samples were genotyped using MALDI-TOF MS analysis.
The SNPs of 1004 Jr (a-) samples were identified using the HRM analysis and DNA sequencing, and 799 of 1004 (80%) Jr (a-) samples had the homozygous for c.376 T. The designed primers for MALDI-TOF MS perfectly detected the SNPs of the ABCG2 gene. A total of 205 Jr (a-) samples were genotyped using MALDI-TOF MS. Calling failures occurred in only two samples with the mutations c.736CT to c.376C and c.421C to c.421CA. The concordance rate between the pre-genotyped and MALDI-TOF MS-based genotyping results was very high (99·02%) for all ABCG2 alleles.
Jr (a-) Japanese donors had almost the homozygous for c.376 T. However, detections of more than 20 SNPs of the ABCG2 gene for the JR blood group genotyping are needed. MALDI-TOF MS-based genotyping was highly concordant with the pre-genotyped results for all ABCG2 alleles.
本研究旨在评估基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）用于ABCG2基因基因分型的适用性，并估计日本人群中的等位基因频率。
Jr(a-)血型在日本献血者中的发生率约为0.05%；除血清学分型外，还进行了基于DNA的基因分型以研究Jr(a-)血型的分子基础。为检测ABCG2基因的所有单核苷酸多态性（SNP），需要一个高通量SNP基因分型平台。
总共从日本献血者中收集了1004份Jr(a-)血样并进行预基因分型。为使用MALDI-TOF MS检测ABCG2基因的SNP，设计了聚合酶链反应和非延伸引物。总共205份Jr(a-)样本通过MALDI-TOF MS分析进行基因分型。
使用高分辨率熔解曲线分析（HRM）和DNA测序鉴定了1004份Jr(a-)样本的SNP，1004份Jr(a-)样本中的799份（80%）为c.376T纯合子。为MALDI-TOF MS设计的引物完美检测到了ABCG2基因的SNP。总共205份Jr(a-)样本通过MALDI-TOF MS进行基因分型。仅在两个发生c.736CT到c.376C和c.421C到c.421CA突变的样本中出现基因分型失败。所有ABCG2等位基因的预基因分型结果与基于MALDI-TOF MS的基因分型结果之间的一致性率非常高（99.02%）。
日本Jr(a-)献血者几乎为c.376T纯合子。然而，JR血型基因分型需要检测ABCG2基因的20多个SNP。基于MALDI-TOF MS的基因分型与所有ABCG2等位基因的预基因分型结果高度一致。
