Zhuo Chuan-Shang, Zhuo Xiao-Fu, Guo Yong-Jian, Wang Chang-Qing
Fujian Province Blood Center, Fuzhou 350004, Fujian Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2008 Apr;16(2):435-8.
To investigate the RHD gene profiles of RhD-negative individuals in population of Fujian Province, it was to design fourteen pairs of specific primers to amplify RHD exon 1, 3 approximately 7, 9, 10, hybrid Rh box, RHD 1227A allele, RHC allele, RHc allele, RHE allele and RHe allele. Rh genotypes were detected by PCR-SSP in 104 RhD-negative donors, some samples with or without RHD genes were analysed by the absorption-elution test, and two RhD-negative samples with eight RHD exons detected were analysed by DNA sequencing. The results showed that 61.54% RhD-negative individuals lacked all the eight RHD exons detected (RHD-/RHD-), 25.97% carried the RHD 1227A allele (62.96% of which were the heterozygote of RHD+/RHD-, and 37.04% were the homozygote of RHD+/RHD+), 8.65% carried the RHD-CE (2 approximately 9)-D allele (RHD+/RHD-), and 1.92% carried the RHD 710delC allele (RHD+/RHD-). Though the most cases of RHD gene deletion were found in dce haplotype, six cases of RHD gene deletion were found in dCe (their RH genotypes were dce/dCe) and two in dcE haplotype (their RH genotypes were dce/dcE). And it was not accurate to predict the Rh phenotype by detecting a single RHD exon, however, and more accurate when eight RHD exons and RHD 1227A allele were detected (chi2=24.43, p<0.005). It is concluded that RHD genes in population of Fujian Province are polymorphic and the RHD genotyping is not reliable enough to replace the RhD serotyping in China.
为研究福建省人群中RhD阴性个体的RHD基因谱,设计了14对特异性引物,用于扩增RHD外显子1、3、约7、9、10、杂交Rh盒、RHD 1227A等位基因、RHC等位基因、RHc等位基因、RHE等位基因和RHe等位基因。采用聚合酶链反应-序列特异性引物(PCR-SSP)检测104例RhD阴性献血者的Rh基因型,部分有或无RHD基因的样本采用吸收-洗脱试验进行分析,对2例检测到8个RHD外显子的RhD阴性样本进行DNA测序。结果显示,61.54%的RhD阴性个体缺乏所有检测的8个RHD外显子(RHD-/RHD-),25.97%携带RHD 1227A等位基因(其中62.96%为RHD+/RHD-杂合子,37.04%为RHD+/RHD+纯合子),8.65%携带RHD-CE(2~9)-D等位基因(RHD+/RHD-),1.92%携带RHD 710delC等位基因(RHD+/RHD-)。虽然RHD基因缺失的大多数病例见于dce单倍型,但在dCe单倍型中发现6例RHD基因缺失(其Rh基因型为dce/dCe),在dcE单倍型中发现2例(其Rh基因型为dce/dcE)。然而,通过检测单个RHD外显子来预测Rh表型并不准确,而检测8个RHD外显子和RHD 1227A等位基因时更准确(χ2=24.43,p<0.005)。结论是福建省人群中RHD基因具有多态性,在中国RHD基因分型可靠性不足,尚不能取代RhD血清学分型。