湖南省RhD阴性献血者D基因多态性研究

Study on RHD Gene Polymorphism Among RhD-negative Blood Donors in Hunan

目的血清学方法研究湖南省133名Rh阴性献血者RHD基因多态性的分布以及RHD基因组结构与血清学的关系。方法应用PCR-SSP技术检测湖南省133名Rh阴性献血者的RHD基因的第2、3、4、5、6、7、9、10外显子、1227A等位基因和RHCE基因的第1、2及5外显子,并分别检测二者的第4内显子。并对部分样本进行吸收放散实验。结果133例Rh阴性个体中,ccee表型69例、Ccee57例、CCee4例、CcEe3例、ccEe1例;133例Rh阴性个体中85例RHD基因所检测的8个外显子完全缺失(63.9%)、24例完整(18.05%)、16例部分缺失(12.03%)。吸收放散试验检测出24例RhD放散型。结论湖南省Rh阴性人群的RHD基因结构呈现多态性,常规血清学鉴定的RhD阴性者中存在较高比例的RhD放散型(Del型),且部分带有完整的RHD基因。携带D基因的吸收放散试验阴性个体中具有C抗原者比例较高,但亦有cc表型存在。提示在中国用RHD基因分型完全替代血清学检测RhD表型条件尚不成熟,仍需二者结合以提高检测的准确性。

Objective To evaluate the distribution of RHD specific exons in 133 blood donors in Hunan Province who were classified as RhD - negative by serological methods, and to explore the association between the genomic structure and serological results. Methods The polymerase chain reaction usingsequence specific primers （PCR - SSP） was used to amplify exons 2, 3, 4, 5, 6, 7, 9 and 10 of RHD gene, allele 1227A and exons 1, 2 and 5 of RHCE gene, as well as intron 4 in each of them. Absorption - elution tests were done in a part of the samples. Results The 133 cases of RhD - negative phenotypes consisted of 69 ccee, 57 Ccee, 4 COee, 3 CcEe, and 1 ccEe. Among them, 85 cases lacked the eight RHD ex- ons indicated above （63.9）, 24 cases had all the RHD exons examined （18.05 % ）, while 16 individuals carried at least one RHD exon （ 12.03 % ）. 24 samples were Del phenotype by the absorption - elution tests. Conclusions Polymorphisms of Rh - negative individuals in Hunan population were observed. Del phenotypes were highly detected among the apparently RhD - negative individuals and Del cases had the possibility of carrying the intact RHD gene. The RhD - negative individuals car- ried RHD gene were mainly found the RHC phenotype while Rhc phenotypes among the RhD - negative cases were also found. So it is necessary to combine serological study with gene phenotyping before DNA- based RhD genotyping is performed.