Mateus-Nino Julio F, Wynn Julia, Wiggins-Smith Jenny, Bryant J Brett, Citty J Kris, Citty J Kyle, Ahuja Samir, Newman Roger
Atrium Health, Concord, North Carolina; BillionToOne Inc, Menlo Park, California; the Medical University of South Carolina, Charleston, South Carolina; Shannon Health, San Angelo, Texas; Unity Health Searcy, Searcy, Arkansas; Wellstar Health System, Marietta, Georgia; and University Hospital, Mentor, Ohio.
Obstet Gynecol. 2025 Apr 1;145(4):402-408. doi: 10.1097/AOG.0000000000005850. Epub 2025 Feb 27.
To evaluate the performance of a cell-free DNA (cfDNA) assay that uses next-generation sequencing with quantitative counting templates for the clinical detection of the fetal RHD genotype in a diverse RhD-negative pregnant population in the United States.
This retrospective cohort study was conducted in four U.S. health care centers. The same next-generation sequencing quantitative counting template cfDNA fetal RhD assay was offered to nonalloimmunized RhD-negative pregnant individuals as part of clinical care. Rh immune globulin (RhIG) was administered at the discretion of the clinician. The sensitivity, specificity, and accuracy of the assay were calculated considering the neonatal RhD serology results.
A total of 401 nonalloimmunized RhD-negative pregnant individuals who received clinical care in the period from August 2020 to November 2023 were included in the analysis. The D antigen cfDNA result was 100% concordant with the neonatal serology, resulting in 100% sensitivity, 100% positive predictive value (95% CI, 98.6-100% for both), 100% specificity, and 100% negative predictive value (95% CI, 97.4-100% for both). There were 10 pregnant individuals in whom the cfDNA analysis identified a non- RHD gene deletion, including RhDΨ (n=5) and RHD-CE-D hybrid variants (n=5). Rh immune globulin was administered antenatally to 93.1% of pregnant individuals, with cfDNA results indicating an RhD-positive fetus compared with 75.0% of pregnant individuals with cfDNA results indicating an RhD-negative fetus, signifying that clinicians were using the cfDNA results to guide pregnancy management.
This next-generation sequencing with quantitative counting templates cfDNA analysis for detecting fetal RhD status is highly accurate with no false-positive or false-negative results in 401 racially and ethnically diverse pregnant individuals with 100% follow-up of all live births. This study and prior studies of this assay support a recommendation to offer cfDNA screening for fetal Rh status as an alternative option to prophylactic RhIG for all nonalloimmunized RhD-negative individuals, which will result in more efficient and targeted prenatal care with administration of RhIG only when medically indicated.
评估一种游离DNA（cfDNA）检测方法的性能，该方法使用下一代测序和定量计数模板，用于在美国不同的RhD阴性孕妇群体中临床检测胎儿RHD基因型。
这项回顾性队列研究在美国的四个医疗保健中心进行。作为临床护理的一部分，为未发生同种免疫的RhD阴性孕妇提供相同的下一代测序定量计数模板cfDNA胎儿RhD检测。Rh免疫球蛋白（RhIG）由临床医生酌情使用。根据新生儿RhD血清学结果计算该检测方法的敏感性、特异性和准确性。
共有401名在2020年8月至2023年11月期间接受临床护理的未发生同种免疫的RhD阴性孕妇被纳入分析。D抗原cfDNA结果与新生儿血清学结果100%一致，敏感性为100%，阳性预测值为100%（两者的95%置信区间均为98.6 - 100%），特异性为100%，阴性预测值为100%（两者的95%置信区间均为97.4 - 100%）。有10名孕妇的cfDNA分析鉴定出非RHD基因缺失，包括RhDΨ（n = 5）和RHD - CE - D杂交变体（n = 5）。cfDNA结果显示为RhD阳性胎儿的孕妇中，93.1%在产前接受了RhIG治疗；cfDNA结果显示为RhD阴性胎儿的孕妇中，这一比例为75.0%，这表明临床医生正在使用cfDNA结果来指导妊娠管理。
这种使用定量计数模板的下一代测序cfDNA分析用于检测胎儿RhD状态的方法高度准确，在401名种族和民族多样化的孕妇中没有假阳性或假阴性结果，并且对所有活产进行了100%的随访。本研究以及此前对该检测方法的研究支持一项建议，即对于所有未发生同种免疫的RhD阴性个体，提供cfDNA筛查胎儿Rh状态作为预防性RhIG的替代选择，这将带来更高效、更有针对性的产前护理，仅在医学指征明确时才使用RhIG。
