OBJECTIVE To present a large U.S. clinical validation of a next-generation sequencing-based, noninvasive prenatal cell-free DNA test for fetal RHD. METHODS This clinical validation study assessed the performance of a commercially available, next-generation sequencing-based cell-free DNA test for fetal RHD status. Samples that passed quality metrics were included if the patient had a previously reported cell-free DNA result for fetal aneuploidy, maternal RhD-negative serology, newborn RhD serology, and maternal RHD deletion or RHD-CE-D hybrid(r's) genotype. Dizygotic twin pregnancies were excluded. Maternal and fetal RHD genotypes were evaluated with prospective cell-free DNA next-generation sequencing analysis. At the time of analysis, investigators were blinded to fetal RhD status. RESULTS The cohort consisted of 655 pregnant patients with serologic results for RhD antigen. Patient demographics included a representative distribution of race and ethnicities in the RhD-negative U.S. population (74.0% White, 13.7% Hispanic, 7.0% Black, and 2.1% Asian). Cell-free DNA fetal RHD was not reported in two cases. There were zero false-negative cases; 356 of 356 fetuses were correctly identified as fetal RhD positive (sensitivity 100%, 95% CI, 98.9-100%). Of the 297 RhD-negative fetuses, 295 were correctly identified as RhD negative (specificity 99.3%, 95% CI, 97.6-99.8%). Of the fetuses with a negative RhD phenotype, the cell-free DNA test accurately identified three with the fetal RHD pseudogene (RHDΨ) genotype. CONCLUSION Validation of this test in this large U.S. cohort of RhD-negative patients provides data on early and accurate noninvasive prenatal identification of fetal RHD genotype at 9 weeks of gestation or more. This test has the potential to assist patients and clinicians in the prevention and management of RhD alloimmunization.

中文翻译：

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在美国大型队列中胎儿 RHD 的产前游离 DNA 筛查测试的临床验证。


目的 介绍用于胎儿 RHD 的下一代基于测序的无创产前游离 DNA 检测的美国大型临床验证。方法 这项临床验证研究评估了市售的、基于下一代测序的胎儿 RHD 状态游离 DNA 检测的性能。如果患者先前报道过胎儿非整倍体、母体 RhD 阴性血清学、新生儿 RhD 血清学和母体 RHD 缺失或 RHD-CE-D 杂交 （r） 基因型的游离 DNA 结果，则包括通过质量指标的样本。排除了异卵双胎妊娠。使用前瞻性游离 DNA 下一代测序分析评估母体和胎儿 RHD 基因型。在分析时，研究人员对胎儿 RhD 状态不知情。结果 该队列由 655 例 RhD 抗原血清学结果的孕妇组成。患者人口统计学包括 RhD 阴性美国人群中种族和民族的代表性分布 （74.0% 白人、13.7% 西班牙裔、7.0% 黑人和 2.1% 亚裔）。2 例未报道胎儿游离 DNA RHD。假阴性病例为零;356 例胎儿中有 356 例被正确鉴定为胎儿 RhD 阳性 （敏感性 100%，95% CI，98.9-100%）。在 297 例 RhD 阴性胎儿中，295 例被正确鉴定为 RhD 阴性 （特异性 99.3%，95% CI，97.6-99.8%）。在 RhD 表型阴性的胎儿中，游离 DNA 测试准确识别了三个具有胎儿 RHD 假基因 （RHDΨ） 基因型的胎儿。结论 在这一大型美国 RhD 阴性患者队列中验证该测试提供了在妊娠 9 周或更长时间时胎儿 RHD 基因型的早期和准确无创产前鉴定的数据。 该测试有可能帮助患者和临床医生预防和管理 RhD 同种异体免疫。