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Fetal RHD genotyping by analysis of maternal plasma in a mixed population

机译:通过混合人群母体血浆分析胎儿RHD基因型

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摘要

: Maternal plasma analysis for the determination of the fetal status is an exciting tool for the management of RhD‐negative pregnant women, specially sensitized women. We assessed the accuracy of fetal genotyping by analysis of maternal plasma in a multi‐ethnic population. : We analyzed plasma samples from 88 RhD‐negative pregnant women between 11 and 39 weeks of gestation, median age of 28 years old to determine the fetal genotype. This population was from Southeastern Brazil with high mixed ethnic background. Fourteen patients (16%) had anti‐D alloantibody. We used Taqman primers and probes to detect by real‐time PCR, exons 4, 5, and 10 of . As internal controls we used primers/probes sets to and . Peripheral or umbilical cord bloods from respective nenonates were collected during delivery and hemagglutination was performed. : Fifty‐eight samples (66%) were genotyped as +, 27 samples (31%) showed complete absence of and 3 samples (3 %) presented a D variant ( ψ). All the results agreed with the neonatal typing, including the three fetuses with the ψ, phenotyped as RhD‐negative. Thus, the accuracy of the fetal genotyping in this mixed population was 100%. The earliest pregnancy in which fetal was detected was 11 weeks. : Our findings indicate that the accuracy of gene using three regions (exons 4, 5, and 10) can be sufficient for clinical application in a multi‐ethnic population. This knowledge helped us on the development of a feasible protocol for fetal genotyping on DNA from maternal plasma for our population. J. Clin. Lab. Anal. 25:100–104, 2011. © 2011 Wiley‐Liss, Inc.
机译::孕妇血浆分析以确定胎儿的状况是管理RhD阴性孕妇(特别是敏感妇女)的令人兴奋的工具。我们通过分析多种族人群的母体血浆评估了胎儿基因分型的准确性。 :我们分析了88名RhD阴性孕妇在11至39周妊娠之间的血浆样本,中位年龄为28岁,以确定胎儿的基因型。该人口来自巴西东南部,具有高度混合的种族背景。 14名患者(16%)患有抗-D同种抗体。我们使用Taqman引物和探针通过实时PCR检测的外显子4、5和10。作为内部对照,我们使用了引物/探针对和。在分娩过程中,从各自的壬酸酯中采集外周血或脐带血,并进行血凝。 :58个样本(66%)的基因型为+,27个样本(31%)显示完全不存在,3个样本(3%)显示为D变体(ψ)。所有结果均与新生儿分型相符,包括三只表型为RhD阴性的ψ胎儿。因此,该混合人群中胎儿基因分型的准确性为100%。最早发现胎儿的妊娠为11周。 :我们的发现表明,使用三个区域(第4、5、10外显子)的基因准确度足以在多族裔人群中进行临床应用。这些知识帮助我们为我们的人群开发了一种可行的方案,用于对来自孕妇血浆的DNA进行胎儿基因分型。 J.临床实验室肛门25:100–104,2011.©2011 Wiley‐Liss,Inc.

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