使用基于荧光的聚合酶链反应对混合样本中的RhD基因型进行灵敏测定。

Sensitive determination of the RhD genotype in mixed samples using fluorescence-based polymerase chain reaction.

作者信息

Tonn T, Westrup D, Seidl C, Kirchmaier C M, Seifried E

机构信息

Institut für Transfusionsmedizin und Immunhämatologie, Blutspendedienst-Hessen des Deutschen Roten Kreuzes, Frankfurt am Main, Deutschland.

出版信息

Vox Sang. 1997;72(3):177-81. doi: 10.1046/j.1423-0410.1997.7230177.x.

DOI:10.1046/j.1423-0410.1997.7230177.x

PMID:9145489

Abstract

OBJECTIVES

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.

METHODS

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.

RESULTS

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.

CONCLUSION

This method will be valuable in prenatal RhD typing after amniocentesis or after the isolation of fetal cells from maternal peripheral blood.

摘要

目的

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。