一种使用游离胎儿 DNA 进行非侵入性胎儿 RHD 基因分型的多重 PCR 方法。

A multiplex PCR for non-invasive fetal RHD genotyping using cell-free fetal DNA.

机构信息

Department of Medical Genetics, Athens University School of Medicine, Athens, Greece.

出版信息

In Vivo. 2011 May-Jun;25(3):411-7.

PMID:21576416

Abstract

AIM

To design a protocol for non-invasive prenatal diagnosis of fetal Rhesus D (RhD) status.

MATERIALS AND METHODS

A total of 112 single lymphocytes were used to test the efficiency of the assay. The protocol was validated using blood samples from 84 RhD-negative pregnant women at 7-24 weeks of gestation. Cell-free DNA (cfDNA) was enzymatically digested using AciI and analyzed by a polymerase chain reaction (PCR) that allowed simultaneous amplification of RHD exons 7 and 10, SRY, RASFF1A and ACTB.

RESULTS

On the one genome-equivalent level, the efficiency of the protocol was ≥ 94.6% for each locus amplified. Conclusive results from the first set of PCRs were obtained for 79 cases with one false-positive. In five cases the analysis was repeated and, subsequently, all cases were accurately diagnosed.

CONCLUSION

The proposed protocol is rapid, applicable in most molecular diagnostic laboratories and provides the basis for non-invasive examination of fetal RhD with 96.7% specificity and 100% sensitivity.

摘要

目的

设计一种用于无创性产前诊断胎儿 RhD（RhD）状态的方案。

材料与方法

共使用 112 个单个淋巴细胞来测试该检测方法的效率。该方案使用了 84 名 RhD 阴性孕妇在妊娠 7-24 周的血液样本进行了验证。使用 AciI 对无细胞 DNA（cfDNA）进行酶切消化，通过聚合酶链反应（PCR）进行分析，该 PCR 允许同时扩增 RHD 外显子 7 和 10、SRY、RASFF1A 和 ACTB。