Standardization non-invasive fetal RHD and SRY determination into clinical routine using a new multiplex RT-PCR assay for fetal cell-free DNA in pregnant women plasma: Results in clinical benefits and cost saving
Standardization non-invasive fetal RHD and SRY determination into clinical routine using a new multiplex RT-PCR assay for fetal cell-free DNA in pregnant women plasma: Results in clinical benefits and cost saving
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DOI：
10.1016/j.cca.2011.11.004
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发表时间：
2012-02-18
影响因子：
5
通讯作者：
Guerrero, Juan M.
中科院分区：
文献类型：
--
作者：
Macher, Hada C.;Noguerol, Pilar;Guerrero, Juan M.
介绍：在阴性RhD母亲中，了解胎儿RhD状态以避免新生儿溶血病的可能性是至关重要的。在这方面，检测孕妇血浆中的胎儿DNA可能成为一种新的诊断工具。在目前的研究中，我们已经评估了标准化的多重PCR针对两个外显子的RHD和一个SRY基因，以监测RhD阴性的妇女。目前的研究解决了有关的可行性和适用性的问题，这种方法到临床practice.Materials和方法：单一和多重实时PCR靶向RHD外显子5和7和SRY被应用于检测胎儿特定的RHD序列和性别在母体血浆。一个大型队列的2127名妇女进行了研究之间的10和28周的怀孕。其中134个用于单TaqMan PCR研究，1993个使用多重TaqMan PCR研究进行评价。根据西班牙指南，所有人的血清学分型均为RhD阴性。单和多重实时PCR结果与出生后血清学和性别鉴定进行了比较。结果：单和多重实时PCR检测结果之间的一致性为100%。目前，1012年的1993名孕妇研究分娩和RHD状态的多重TaqMan PCR检测获得的结果证实产后血清学方法显示，敏感性，特异性和准确性的多重检测分别为100，98.6和99.3%，分别。该方法提高了检测速度，避免了RhD阴性孕妇携带RhD阴性胎儿时的过度治疗，降低了临床和生物学监测的要求，具有较好的临床效益和成本节约。结论：应用多重实时PCR技术，常规检测孕妇血浆中胎儿RHD状态和SRY是可行的。多重实时PCR的使用可以提高实验室的响应，节省时间和试剂成本，为该过程的完全自动化打开大门。(C)2011 Elsevier B.V.保留所有权利。
Introduction: Among negative RhD mothers it is essential to know the fetal RhD status in order to avoid the possibility of hemolytic disease of the newborn. In this regard, the detection of fetal DNA in maternal plasma might become a new diagnostic tool. In the current study, we have evaluated the standardization of a Multiplex-PCR targeted towards two exons of the RHD and one SRY gene to monitor RhD negative women. The current study addresses questions concerning feasibility and applicability of this approach into the clinical practice.Materials and methods: Both single and multiplex real-time PCRs targeting RHD exons 5 and 7 and SRY were applied for the detection of fetal-specific RHD sequences and sex in maternal plasma. A large cohort of 2127 women was studied between 10 and 28 weeks of pregnancy. 134 of them were used for single TaqMan PCR studies and 1993 were evaluated using Multiplex TaqMan PCR studies. All of them were serologically typed as RhD negative according to Spanish guidelines. Single and multiplex real-time PCR results were compared with postnatal serology and sex identification.Results: There was a 100% concordance between results obtained with single and multiplex real-time PCR assays. At present, 1012 of the 1993 pregnant women studied gave birth and the results of RHD status obtained with the multiplex TaqMan PCR assay were confirmed postpartum by serological methods showing that sensitivity, specificity, and accuracy of the multiplex assay were 100, 98.6, and 99.3%, respectively. This procedure improved the speed of the assay, avoided over-treatment among RhD negative pregnant women bearing RhD negative fetus, and reduced the requirements for clinical and biological monitoring, resulting in a clinical benefit and cost saving.Conclusions: The routine determination of fetal RHD status and SRY in maternal plasma, using multiplex real-time PCR, is feasible. The use of multiplex real-time PCR allows improving the response of the laboratory, saving time and reagent costs, opening the door to a complete automatization of the process. (C) 2011 Elsevier B.V. All rights reserved.