Clausen Frederik Banch, Barrett Angela Natalie
Department of Clinical Immunology, Rigshospitalet, Copenhagen University Hospital, Copenhagen, Denmark.
Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore City, Singapore.
Vox Sang. 2019 May;114(4):386-393. doi: 10.1111/vox.12768. Epub 2019 Mar 4.
Fetal RHD genotyping of cell-free fetal DNA from RhD-negative pregnant women can be used to guide targeted antenatal and postnatal anti-D prophylaxis for the prevention of RhD immunization. To assure the quality of clinical testing, we conducted an external quality assessment workshop with the participation of 28 laboratories.
Aliquots of pooled maternal plasma were sent to each laboratory. One sample was positive, and the second sample was negative for fetal RHD, verified by pre-workshop testing using quantitative real-time PCR (qPCR) analysis of RHD exons 4, 5, 7 and 10. Plasma samples were shipped at room temperature. A reporting scheme was supplied for data collection, including questions regarding the methodological setup, results and clinical recommendations. Different methodological approaches were used, all employing qPCR with a total of eight different combinations of RHD exon targets. The samples were tested blindly.
Fetal RHD genotyping was performed with no false-negative and no false-positive results. One inconclusive result was reported for the RHD-positive sample, and four inconclusive results were reported for the RHD-negative sample. All clinical conclusions were satisfactory.
This external quality assessment workshop demonstrates that despite the different approaches taken to perform the clinical assays, fetal RHD genotyping is a reliable laboratory assay to guide targeted use of Rh prophylaxis in a clinical setting.
对RhD阴性孕妇的游离胎儿DNA进行胎儿RHD基因分型，可用于指导有针对性的产前和产后抗D预防，以防止RhD免疫。为确保临床检测质量，我们举办了一次有28个实验室参与的外部质量评估研讨会。
将混合的母血浆等分试样发送至每个实验室。通过使用RHD外显子4、5、7和10的定量实时PCR（qPCR）分析进行研讨会前检测，验证其中一个样本的胎儿RHD为阳性，另一个样本为阴性。血浆样本在室温下运送。提供了一个报告方案用于数据收集，包括有关方法设置、结果和临床建议的问题。使用了不同的方法，均采用qPCR，共有八种不同的RHD外显子靶标组合。样本进行盲法检测。
胎儿RHD基因分型未出现假阴性和假阳性结果。RHD阳性样本报告了一个不确定结果，RHD阴性样本报告了四个不确定结果。所有临床结论均令人满意。
本次外部质量评估研讨会表明，尽管临床检测采用了不同方法，但胎儿RHD基因分型是一种可靠的实验室检测方法，可用于指导临床环境中Rh预防的靶向应用。
