采用荧光双重聚合酶链反应对同种免疫妊娠羊水中的胎儿恒河猴D基因型进行检测。

Fetal Rhesus D genotyping on amniocytes in alloimmunised pregnancies using fluorescence duplex polymerase chain reaction.

作者信息

Crombach G, Picard F, Beckmann M, Tutschek B, Bald R, Niederacher D

机构信息

Department of Prenatal Diagnosis, Universities of Düsseldorf, Köln, Germany.

出版信息

Br J Obstet Gynaecol. 1997 Jan;104(1):15-9. doi: 10.1111/j.1471-0528.1997.tb10641.x.

DOI:10.1111/j.1471-0528.1997.tb10641.x

Abstract

OBJECTIVE

To establish the reliability of fetal amniocyte Rhesus D (RhD) genotyping using fluorescence duplex polymerase chain reaction (PCR) and 2. to assess the potential clinical impact on management of alloimmunised pregnancies.

DESIGN

Multicentre observational study.

SETTING

Four departments of obstetrics and gynaecology in Germany.

METHODS

Fourty-four amniotic fluid samples were obtained by amniocentesis from a retrospective group of 27 RhD alloimmunised pregnancies and 15 samples from 14 women treated prospectively. Two RhD gene specific fragments (exon 7 and 10) were amplified using two separate fluorescence duplex PCR assays, and laser detected in an automated DNA sequence analyser.

RESULTS

Amplification of the Rh gene sequences was successful in all samples. PCR at the two RhD gene regions resulted in complete concordance. Genotyping correctly predicted the RhD status of all fetuses serotyped (n = 41). After intrauterine transfusions, PCR identified the RhD type of two fetuses more accurately than serotyping. Earlier knowledge of a negative RhD status would have rendered unnecessary 12 amniocenteses in four fetuses of the retrospective study group, and prevented further invasive testing in one fetus treated prospectively. In the latter group, women with a positive fetal RhD genotype underwent intensive prenatal care including serial invasive monitoring and intrauterine treatment.

CONCLUSIONS

Fetal RhD genotyping of amniocytes is a reliable technique with the potential to improve routine management of alloimmunised pregnant women.

摘要

目的

利用荧光双重聚合酶链反应（PCR）建立胎儿羊膜细胞恒河猴D（RhD）基因分型的可靠性，2. 评估对同种免疫妊娠管理的潜在临床影响。

设计

多中心观察性研究。

地点

德国四个妇产科部门。

方法

通过羊膜穿刺术从27例RhD同种免疫妊娠的回顾性队列中获取44份羊水样本，从14例前瞻性治疗的女性中获取15份样本。使用两种单独的荧光双重PCR检测法扩增两个RhD基因特异性片段（外显子7和10），并在自动DNA序列分析仪中进行激光检测。

结果

所有样本中Rh基因序列的扩增均成功。两个RhD基因区域的PCR结果完全一致。基因分型正确预测了所有进行血清分型的胎儿（n = 41）的RhD状态。在进行宫内输血后，PCR比血清分型更准确地鉴定了两个胎儿的RhD类型。更早知晓RhD阴性状态可使回顾性研究组中四个胎儿的12次羊膜穿刺术不必要，并避免了对一名前瞻性治疗胎儿的进一步侵入性检测。在后一组中，胎儿RhD基因型为阳性的女性接受了强化产前护理，包括系列侵入性监测和宫内治疗。

结论

羊膜细胞胎儿RhD基因分型是一种可靠的技术，有可能改善同种免疫孕妇的常规管理。