目的 建立Junior血型基因分型技术，用于Jr(a-)稀有血型的鉴定和筛查。 方法 选取2021年1月至2021年5月于深圳市血液中心无偿献血的O型RhD＋健康受试者(n＝1 568)和1个疑难交叉配血家系(n＝3)，共1 571例，为研究对象。用血清学检测技术进行先证者血型鉴定、意外抗体鉴定以及抗体效价测定。用聚合酶链反应-序列特异性引物(PCR-SSP)技术进行先证者RHD基因分型。建立ABCG2基因编码区测序和PCR-SSP基因分型技术，对先证者及其家系成员进行基因型检测，并在深圳地区无偿献血者人群(n＝1 568)中开展Jra抗原阴性的稀有血型献血者筛查。 结果 先证者ABO血型为B型，RhD血型为部分D(RHD*DVI.3/RHD*01N.01)，Junior血型Jra抗原为阴性，血浆存在抗-D合并抗-Jra。ABCG2基因测序发现先证者等位基因型为ABGG2*01N.01/ABGG2*01N.01[c.376C>T(p.Gln126X)纯合变异]，为亚洲人群中最常见的Jr(a-)血型等位基因。在深圳地区无偿献血者人群中进行筛查，无Jr(a-)稀有血型献血者检出。通过杂合子的统计分析，发现ABCG2*01N.01(c.376T)的等位基因型频率约为0.45%，这一分子背景的Jr(a-)稀有血型在深圳地区的出现频率约为0.2‰。 结论 本研究发现国内首例部分DVI.3型且Jr(a-)稀有血型，并成功建立Junior血型ABCG2基因编码区测序以及PCR-SSP基因分型技术。 Objective To develop a genotyping method for the Junior blood type and report on a rare blood type with Jr(a-). Methods Healthy O-type RhD+ volunteer donors of the Shenzhen Blood Center from January to May 2021 (n=1 568) and a pedigree with difficult cross-matching (n=3) were selected as the study subjects. Serological methods were used for proband′s blood type identification, unexpected antibody identification, and antibody titer determination. Polymerase chain reaction-sequence specific primer (PCR-SSP) method was used for typing the proband′s RHD gene. ABCG2 gene coding region sequencing and a PCR-SSP genotyping method were established for determining the genotypes of the proband and his family members and screening of Jra antigen-negative rare blood type among the 1 568 blood donors. Results The proband′s ABO and RhD blood types were respectively determined as B and partial D (RHDDVI.3/RHD01N.01), Junior blood type Jra antigen was negative, and plasma had contained anti-D and anti-Jra. Sequencing of the ABCG2 gene revealed that the proband′s genotype was ABGG201N.01/ABGG201N.01 [homozygous c. 376C>T (p.Gln126X) variants], which is the most common Jr(a-) blood type allele in the Asian population. Screening of the voluntary blood donors has detected no Jr(a-) rare blood type. Statistical analysis of the heterozygotes suggested that the allelic frequency forABCG2*01N.01 (c.376T) was 0.45%, and the frequency of Jr(a-) rare blood type with this molecular background was about 0.2‰. Conclusion A very rare case of partial DVI.3 type and Jr(a-) rare blood type has been identified. And a method for identifying the Junior blood type through sequencing the coding regions of the ABCG2 gene and PCR-SSP has been established.
Establishment of a genotyping method for the junior blood group and identification of a rare blood type with partial DVI.3 and Jr(a-)
Objective To develop a genotyping method for the Junior blood type and report on a rare blood type with Jr(a-). Methods Healthy O-type RhD+ volunteer donors of the Shenzhen Blood Center from January to May 2021 (n=1 568) and a pedigree with difficult cross-matching (n=3) were selected as the study subjects. Serological methods were used for proband′s blood type identification, unexpected antibody identification, and antibody titer determination. Polymerase chain reaction-sequence specific primer (PCR-SSP) method was used for typing the proband′s RHD gene. ABCG2 gene coding region sequencing and a PCR-SSP genotyping method were established for determining the genotypes of the proband and his family members and screening of Jra antigen-negative rare blood type among the 1 568 blood donors. Results The proband′s ABO and RhD blood types were respectively determined as B and partial D (RHDDVI.3/RHD01N.01), Junior blood type Jra antigen was negative, and plasma had contained anti-D and anti-Jra. Sequencing of the ABCG2 gene revealed that the proband′s genotype was ABGG201N.01/ABGG201N.01 [homozygous c. 376C>T (p.Gln126X) variants], which is the most common Jr(a-) blood type allele in the Asian population. Screening of the voluntary blood donors has detected no Jr(a-) rare blood type. Statistical analysis of the heterozygotes suggested that the allelic frequency forABCG2*01N.01 (c.376T) was 0.45%, and the frequency of Jr(a-) rare blood type with this molecular background was about 0.2‰. Conclusion A very rare case of partial DVI.3 type and Jr(a-) rare blood type has been identified. And a method for identifying the Junior blood type through sequencing the coding regions of the ABCG2 gene and PCR-SSP has been established.
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