Rouillac-Le Sciellour C, Sérazin V, Brossard Y, Oudin O, Le Van Kim C, Colin Y, Guidicelli Y, Menu M, Cartron J-P
Centre hospitalier intercommunal, Poissy-St-Germain, France.
Transfus Clin Biol. 2007 Dec;14(6):572-7. doi: 10.1016/j.tracli.2008.01.003. Epub 2008 Mar 28.
Fetal RHD genotyping from maternal plasma was performed by real-time PCR amplification of exons 7 and 10 of the RHD gene and the amplified products were detected either with SYBR Green I dye according to our previously published method [Mol Diagn 8 (2004) 23-31] or with hydrolysis probes in a new Free DNA Fetal Kit RhD((R)). Plasma specimen from 300 RhD-negative pregnant women (between 10 to 34 weeks of gestation) were analysed and validation of the results was ascertained either by RHD genotyping on amniotic cells or by blood typing of the neonate at birth. We found 100% concordant results when comparing the two methods. Two false-positive but no false-negative results were found. Thus, the sensitivity of the assay was 100% and the specificity superior than 99%. These data confirm the accuracy of fetal RHD genotyping on maternal plasma using the Free DNA Fetal Kit RhD, thus allowing to propose non invasive PCR-based fetal RHD genotyping for all RhD-negative pregnant women and to restrict the use of anti-D immunoglobulins only to those bearing an RhD-positive fetus.
通过对RHD基因外显子7和10进行实时PCR扩增，从孕妇血浆中进行胎儿RHD基因分型，并根据我们之前发表的方法[《分子诊断》8(2004)23 - 31]，用SYBR Green I染料或在新型游离DNA胎儿RhD检测试剂盒中用水解探针检测扩增产物。分析了300名RhD阴性孕妇（妊娠10至34周）的血浆样本，并通过对羊水细胞进行RHD基因分型或在新生儿出生时进行血型鉴定来确定结果的有效性。比较这两种方法时，我们发现结果100%一致。发现了2例假阳性结果，但无假阴性结果。因此，该检测方法的灵敏度为100%，特异性高于99%。这些数据证实了使用游离DNA胎儿RhD检测试剂盒对孕妇血浆进行胎儿RHD基因分型的准确性，从而可以为所有RhD阴性孕妇提供基于非侵入性PCR的胎儿RHD基因分型，并仅将抗D免疫球蛋白的使用限制在怀有RhD阳性胎儿的孕妇中。
