Junior血型基因检测技术的建立与1例部分DVI.3型且Jr(a-)稀有血型的鉴定

Establishment of a genotyping method for the junior blood group and identification of a rare blood type with partial DVI.3 and Jr(a-)

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=1568)和1个疑难交叉配血家系(n=3),共1571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=1568)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=1568)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 1568 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 for ABCG2*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.