Maas J H, Legler T J, Lynen R, Blaschke V, Ohto H, Köhler M
Abteilung Transfusionsmedizin, Universität Göttingen, Deutschland.
Beitr Infusionsther Transfusionsmed. 1997;34:203-9.
RHD genotyping from fetal cells was applied for the detection of the RHD gene in the fetus of immunized Rh-D-negative women. Additionally, RHD genotyping was applied for the characterization of Rh-D variants. Although 44 nucleotide substitutions are known to code for 35 amino acid differences between the RHCE and the RHD gene, only a few polymorphisms have been investigated yet. We investigated 7 RHD-specific nucleotides on exons 2, 5, and 7 with sequence-specific primers and 1 nucleotide with ligation-based typing. All RHD genotyping results were correlated with serological results and established genotyping methods in 116 German and 98 Japanese blood donors, because different genetic sequences coding for Rh-D polypeptides have been described in different ethnic groups. Sequence-specific amplification of D-specific sequences was concordant with the serological result in all blood donors tested. However, ligation-based typing on exon 5 gave false-negative results in 7 donors. In summary, 5 new sequence-specific PCRs have been evaluated for further characterization of Rh-D variants. Furthermore, the methods described allow nested PCR and thus may help in determination of the fetal RhD status from maternal peripheral blood during pregnancy.
对免疫的Rh-D阴性女性胎儿的细胞进行RHD基因分型，以检测胎儿中的RHD基因。此外，RHD基因分型还用于Rh-D变异体的特征分析。虽然已知有44个核苷酸替换可编码RHCE和RHD基因之间35个氨基酸的差异，但目前仅研究了少数多态性。我们使用序列特异性引物研究了外显子2、5和7上的7个RHD特异性核苷酸，并使用基于连接的分型方法研究了1个核苷酸。由于不同种族群体中已描述了编码Rh-D多肽的不同基因序列，因此我们将所有RHD基因分型结果与116名德国和98名日本献血者的血清学结果及既定基因分型方法进行了关联。在所有检测的献血者中，D特异性序列的序列特异性扩增与血清学结果一致。然而，外显子5上基于连接的分型在7名献血者中给出了假阴性结果。总之，已评估了5种新的序列特异性PCR，以进一步表征Rh-D变异体。此外，所描述的方法允许进行巢式PCR，因此可能有助于在孕期从母体外周血中确定胎儿的RhD状态。
