Tonn T, Westrup D, Seidl C, Kirchmaier C M, Seifried E
Institut für Transfusionsmedizin und Immunhämatologie, Blutspendedienst-Hessen des Deutschen Roten Kreuzes, Frankfurt am Main, Deutschland.
Vox Sang. 1997;72(3):177-81. doi: 10.1046/j.1423-0410.1997.7230177.x.
Prenatal determination of the fetal RhD status in pregnancies of Rh-negative (Rh-neg) women by polymerase chain reaction (PCR) on DNA has become of increasing importance. Since determination of the RhD genotype in these cases is usually performed in samples containing both maternal Rh-neg and fetal Rh-neg or Rh-positive (Rh-pos) DNA, the sensitivity and specificity of PCR-based DNA detection are of crucial importance in the diagnosis of the fetal RhD status.
We developed a method for RhD typing using the PCR and a fluorescence-based detection method that allows us to determine RhD-pos DNA even if it is mixed with large amounts of Rh-neg DNA.
Using this approach, Rh-pos DNA could be detected in dilutions with Rh-neg DNA of as high as 1 in 10,000 (Rh-pos/Rh-neg). Furthermore PCR amplification could also be carried out on DNA samples from persons with weak (Dweak) or partial (Dcat) RhD antigens.
This method will be valuable in prenatal RhD typing after amniocentesis or after the isolation of fetal cells from maternal peripheral blood.
通过对DNA进行聚合酶链反应（PCR）来产前确定Rh阴性（Rh-neg）孕妇的胎儿RhD状态已变得越来越重要。由于这些病例中RhD基因型的测定通常在同时含有母体Rh-neg和胎儿Rh-neg或Rh阳性（Rh-pos）DNA的样本中进行，基于PCR的DNA检测的灵敏度和特异性对于胎儿RhD状态的诊断至关重要。
我们开发了一种使用PCR和基于荧光的检测方法进行RhD分型的方法，该方法使我们即使在Rh-pos DNA与大量Rh-neg DNA混合的情况下也能确定Rh-pos DNA。
使用这种方法，在Rh-neg DNA稀释比例高达万分之一（Rh-pos/Rh-neg）的情况下仍可检测到Rh-pos DNA。此外，对具有弱（Dweak）或部分（Dcat）RhD抗原的人的DNA样本也可进行PCR扩增。
该方法对于羊膜穿刺术后或从母体外周血中分离出胎儿细胞后的产前RhD分型将具有重要价值。
