Clinical Pathology Department, Faculty of Medicine, Dewan Aam El Mohafza, Fayoum, Egypt.
Transfusion. 2013 Nov;53(11 Suppl 2):2940-4. doi: 10.1111/trf.12100. Epub 2013 Jan 30.
Rh discrepancies are a problem during routine testing because of partial and weak D phenotypes. Some blood units with weak and partial D expression may escape detection by serology. Limitations of serology can be overcome by molecular typing. The objective of study was to compare currently used serologic methods with molecular analysis to determine the potential application of molecular methods to improve D typing strategies and to estimate the frequency of weak D types among the Arab population.
Fifty blood donor and patient samples with discrepant results of D phenotyping were subjected to routine serology to define the D phenotype including monoclonal anti-D immunoglobulin M and indirect antiglobulin test. Commercially available panels of monoclonal anti-D were used for identification of partial D and weak D phenotypes. Genomic DNA was evaluated using allele-specific amplification polymerase chain reaction with sequence-specific primers to define weak D type.
Molecular typing confirmed most of the serology results; three samples that were not clear-cut serologically were identified by molecular typing, two samples as weak D Type 4.2 (DAR), and one sample as weak D Type 4.0. Another two samples identified by serologic panel as weak D were unresolved by molecular typing. A sample with partial D Type II by serology revealed a Weak D Type 4.0 by molecular typing. Results interestingly showed the high frequency of weak D Type 4.2 (DAR) in Egypt.
RHD molecular typing can solve discrepancies during routine testing due to partial and weak D phenotypes for better transfusion outcome.
由于部分和弱 D 表型,Rh 差异在常规检测中是一个问题。一些具有弱和部分 D 表达的血液单位可能会逃避血清学检测。分子分型可以克服血清学的局限性。本研究的目的是将目前使用的血清学方法与分子分析进行比较,以确定分子方法在改进 D 型检测策略方面的潜在应用,并估计弱 D 型在阿拉伯人群中的频率。
50 份献血者和患者样本的 D 表型检测结果不一致,进行常规血清学检测以确定 D 表型,包括单克隆抗-D 免疫球蛋白 M 和间接抗球蛋白试验。使用商业上可获得的单克隆抗-D 面板来鉴定部分 D 和弱 D 表型。使用等位基因特异性扩增聚合酶链反应和序列特异性引物对基因组 DNA 进行评估,以定义弱 D 型。
分子分型证实了大多数血清学结果;通过分子分型鉴定了三个血清学结果不明确的样本,两个样本为弱 D 型 4.2(DAR),一个样本为弱 D 型 4.0。另外两个通过血清学检测板鉴定为弱 D 的样本无法通过分子分型解决。一个通过血清学鉴定为部分 D 型 II 的样本通过分子分型显示为弱 D 型 4.0。结果显示,埃及弱 D 型 4.2(DAR)的频率很高。
RHD 分子分型可解决由于部分和弱 D 表型导致的常规检测中的差异,以获得更好的输血效果。