Kacem Narjes, Chakroun Tahar, Moussa Hajer, Abdelkefi Saïda, Houissa Betoul, Chiaroni Jacques, Jemni Yacoub Saloua
Centre Régional de Transfusion Sanguine, Unité de recherche UR06SP05, Sousse, Tunisia.
Transfus Med. 2012 Oct;22(5):362-6. doi: 10.1111/j.1365-3148.2012.01172.x. Epub 2012 Jul 4.
Determination of the RHD zygosity is important for genetic counselling and risk evaluation of hemolytic disease of the newborn HDN in women with D iso-immunisation.
We proposed to determine the genotype frequencies of the RHD locus using a PCR-SSP method and assignment of the most probable genotype (MPG) and analyse the concordance between the two methods.
The complete Rh phenotype and the frequencies of RH haplotypes were determined on 506 blood donors. RHD zygosity was determined by both assignment of the MPG and PCR-SSP specific for the hybrid Rhesus box. For RH:-1 samples, analysis of the RHD exon 10 was done to detect eventual RHD aberrant alleles.
Among the 466 RH:1 samples, 54.08% were hemizygous and 45.92% homozygous by PCR-SSP, and 64.16% hemizygous and 35.84% homozygous by the MPG. The comparison between the methods showed discordant results in 135 RH:1 samples. For the 40 RH:-1 samples, hybrid Rhesus box was detected in all samples and RHD exon 10 was detected in three samples suggesting unequivocal alleles identified as one RHDψ, one (C)ce(s) and one weak D type 4.
The PCR-SSP should replace the MPG. However, studying of aberrant RHD alleles and aberrant Rhesus boxes could confirm the accuracy of this method in Tunisian population.
对于D型同种免疫的女性，确定RHD基因的纯合性对于新生儿溶血病（HDN）的遗传咨询和风险评估很重要。
我们提议使用聚合酶链反应-序列特异性引物（PCR-SSP）方法确定RHD基因座的基因型频率，并确定最可能的基因型（MPG），并分析这两种方法之间的一致性。
对506名献血者进行了完整的Rh血型表型和Rh单倍型频率测定。通过MPG的确定和针对杂交恒河猴盒的PCR-SSP来确定RHD基因的纯合性。对于Rh:-1样本，对RHD外显子10进行分析以检测可能存在的RHD异常等位基因。
在466例Rh:1样本中，通过PCR-SSP检测，54.08%为半合子，45.92%为纯合子；通过MPG检测，64.16%为半合子，35.84%为纯合子。两种方法的比较显示，在135例Rh:1样本中结果不一致。对于40例Rh:-1样本，在所有样本中均检测到杂交恒河猴盒，在3个样本中检测到RHD外显子10，提示明确鉴定出一个RHDψ、一个（C）ce（s）和一个弱D 4型等位基因。
PCR-SSP应取代MPG。然而，对异常RHD等位基因和异常恒河猴盒的研究可以证实该方法在突尼斯人群中的准确性。
