Ferro Marta, Macher Hada C, Fornés Gema, Martín-Sánchez Jesús, Jimenez-Arriscado Pilar, Molinero Patrocinio, Pérez-Simón José A, Guerrero Juan M, Rubio Amalia
Instituto de Investigaciones Biomédicas de Sevilla, IBIS (Universidad de Sevilla, HUVR, Junta de Andalucía, CSIC), Hospital Universitario Virgen del Rocío-Universidad de Sevilla, Seville, Andalucía, Spain.
the Blood Transfusion Center and Tissue and Cells Establishment Córdoba, Cordoba, Andalucía, Spain.
Transfusion. 2018 Oct;58(10):2272-2279. doi: 10.1111/trf.14837. Epub 2018 Sep 17.
The development of new noninvasive approaches for the diagnosis of human platelet antigen (HPA)-1 fetomaternal incompatibility has become of great interest. These approaches allow determination of whether the fetus is incompatible or not with the mother and a decision on antenatal therapy to avoid fetal or neonatal alloimmune thrombocytopenia (FNAIT). The objective of this work was to perform rapid, noninvasive prenatal test for HPA-1ab fetal antigen detection after the detection of an HPA-1-homozygous mother by using plasma cell-free DNA (cfDNA).
The HPA-1 genotypes of 142 pregnant women and 17 nonpregnant controls were retrospectively determined by high-resolution melting (HRM) polymerase chain reaction (PCR). Coamplification at lower denaturation temperature (COLD) HRM PCR was performed to determine the fetal genotype analyzing cfDNA from all HPA-1bb pregnant women.
After the HRM analysis, the following genotypes were identified: HPA-1aa (71.13%), HPA-1bb (2.8%), and HPA-1ab (26.06%). Four HPA-1bb-homozygous pregnant women were carrying an incompatible fetus. Plasma samples from these mothers were analyzed by HRM COLD-PCR. Homozygous HPA-1bb pregnant women carrying an HPA-1ab-heterozygous fetus did not group with either the HPA-1ab or the HPA-1bb controls. Thus, COLD-PCR analysis allows the detection of HPA-1ab-heterozygous fetuses carried by homozygous mothers during first weeks of pregnancy.
The fetal genotype from HPA-1bb-homozygous women was detected by a noninvasive prenatal test as soon as 12 weeks of gestation.
开发用于诊断人类血小板抗原（HPA）-1母婴不相容性的新型非侵入性方法已引起广泛关注。这些方法能够确定胎儿与母亲是否不相容，并决定是否进行产前治疗以避免胎儿或新生儿同种免疫性血小板减少症（FNAIT）。本研究的目的是在检测到HPA-1纯合子母亲后，通过使用血浆游离DNA（cfDNA）对HPA-1ab胎儿抗原进行快速、非侵入性产前检测。
通过高分辨率熔解（HRM）聚合酶链反应（PCR）对142名孕妇和17名非孕妇对照的HPA-1基因型进行回顾性测定。采用低温共扩增（COLD）HRM PCR分析所有HPA-1bb孕妇的cfDNA，以确定胎儿基因型。
HRM分析后，鉴定出以下基因型：HPA-1aa（71.13%）、HPA-1bb（2.8%）和HPA-1ab（26.06%）。4名HPA-1bb纯合子孕妇怀有不相容胎儿。通过HRM COLD-PCR对这些母亲的血浆样本进行分析。怀有HPA-1ab杂合子胎儿的HPA-1bb纯合子孕妇既不与HPA-1ab对照组也不与HPA-1bb对照组归为一组。因此，COLD-PCR分析可在妊娠早期检测出纯合子母亲所怀的HPA-1ab杂合子胎儿。
通过非侵入性产前检测，在妊娠12周时即可检测出HPA-1bb纯合子女性的胎儿基因型。
