Ouchari Mouna, Jemni-Yaacoub Saloua, Chakroun Taher, Abdelkefi Saida, Houissa Batoul, Hmida Slama
Unité de Recherche « UR06SP05 » Centre Régional de Transfusion Sanguine, Sousse-Tunisia.
Asian J Transfus Sci. 2013 Jul;7(2):119-24. doi: 10.4103/0973-6247.115568.
A comprehensive survey of RHD alleles in Tunisia population was lacking. The aim of this study was to use a multiplex RHD typing assay for simultaneous detection of partial D especially with RHD/RHCE deoxyribonucleic acid (DNA) sequence exchange mechanism and some weak D alleles.
Six RHD specific primer sets were designed to amplify RHD exons 3, 4, 5, 6, 7 and 9. DNA from 2000 blood donors (1777 D+ and 223 D-) from several regions was selected for RHD genotyping using a PCR multiplex assay. Further molecular investigations were done to characterize the RHD variants that were identified by the PCR multiplex assay.
In the 1777 D+ samples, only 10 individuals showed the absence of amplification of exons 4 and 5 that were subsequently identified by PCR-SSP as weak D type 4 variants. No hybrid allele was detected. In the 223 D-, RHD amplification of some exons was observed only in 5 samples: 4 individuals expressed only RHD exon 9, and one subject lacking exons 4 and 5. These samples were then screened by PCR-SSPs on d(C) ce(s) and weak D type 4, respectively.
The weak D type 4 appears to be the most common D variant allele. We have not found any partial D variant. Findings also indicated that RHD gene deletion is the most prevalent cause of the D- phenotype in the Tunisian population.
突尼斯人群中缺乏对RHD等位基因的全面调查。本研究的目的是使用多重RHD分型检测法同时检测部分D,尤其是具有RHD/RHCE脱氧核糖核酸(DNA)序列交换机制的部分D以及一些弱D等位基因。
设计了6套RHD特异性引物对,用于扩增RHD基因外显子3、4、5、6、7和9。从几个地区的2000名献血者(1777名D阳性和223名D阴性)中选取DNA,使用聚合酶链反应(PCR)多重检测法进行RHD基因分型。对通过PCR多重检测法鉴定出的RHD变异体进行了进一步的分子研究。
在1777份D阳性样本中,只有10人显示外显子4和5无扩增,随后通过PCR-序列特异性引物(PCR-SSP)鉴定为弱D4型变异体。未检测到杂交等位基因。在223份D阴性样本中,仅在5份样本中观察到一些外显子的RHD扩增:4人仅表达RHD外显子9,1名受试者缺乏外显子4和5。然后分别通过PCR-SSP对这些样本进行d(C)ce(s)和弱D4型筛查。
弱D4型似乎是最常见的D变异等位基因。我们未发现任何部分D变异体。研究结果还表明,RHD基因缺失是突尼斯人群中D阴性表型的最普遍原因。