当前位置: X-MOL 学术Transfus. Med. Hemother. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Next Generation Sequencing-Based Fetal ABO Blood Group Prediction by Analysis of Cell-Free DNA from Maternal Plasma
Transfusion Medicine and Hemotherapy ( IF 1.9 ) Pub Date : 2020-01-01 , DOI: 10.1159/000505464
Klaus Rieneck 1 , Christoffer Egeberg Hother 1 , Frederik Banch Clausen 1 , Marianne Antonius Jakobsen 2 , Thomas Bergholt 3 , Ellinor Hellmuth 3 , Lene Grønbeck 3 , Morten Hanefeld Dziegiel 1
Affiliation  

Introduction: ABO blood group incompatibility between a pregnant woman and her fetus as a cause of morbidity or mortality of the fetus or newborn remains an important, albeit rare, risk. When a pregnant woman has a high level of anti-A or anti-B IgG antibodies, the child may be at risk for hemolytic disease of the fetus and newborn (HDFN). Performing a direct prenatal determination of the fetal ABO blood group can provide valuable clinical information. Objective: Here, we report a next generation sequencing (NGS)-based assay for predicting the prenatal ABO blood group. Materials and Methods: A total of 26 plasma samples from 26 pregnant women were tested from gestational weeks 12 to 35. Of these samples, 20 were clinical samples and 6 were test samples. Extracted cell-free DNA was PCR-amplified using 2 primer sets followed by NGS. NGS data were analyzed by 2 different methods, FASTQ analysis and a grep search, to ensure robust results. The fetal ABO prediction was compared with the known serological infant ABO type, which was available for 19 samples. Results: There was concordance for 19 of 19 predictable samples where the phenotype information was available and when the analysis was done by the 2 methods. For immunized pregnant women (n = 20), the risk of HDFN was predicted for 12 fetuses, and no risk was predicted for 7 fetuses; one result of the clinical samples was indeterminable. Cloning and sequencing revealed a novel variant harboring the same single nucleotide variations as ABO*O.01.24 with an additional c.220C>T substitution. An additional indeterminable result was found among the 6 test samples and was caused by maternal heterozygosity. The 2 indeterminable samples demonstrated limitations to the assay due to hybrid ABO genes or maternal heterozygosity. Conclusions: We pioneered an NGS-based fetal ABO prediction assay based on a cell-free DNA analysis from maternal plasma and demonstrated its application in a small number of samples. Based on the calculations of variant frequencies and ABO*O.01/ABO*O.02 heterozygote frequency, we estimate that we can assign a reliable fetal ABO type in approximately 95% of the forthcoming clinical samples of type O pregnant women. Despite the vast genetic variations underlying the ABO blood groups, many variants are rare, and prenatal ABO prediction is possible and adds valuable early information for the prevention of ABO HDFN.

中文翻译:

通过分析母体血浆中的游离 DNA 来预测下一代基于测序的胎儿 ABO 血型

简介: 孕妇与其胎儿之间的 ABO 血型不相容是导致胎儿或新生儿发病或死亡的一个重要风险,尽管这种风险很少见。当孕妇具有高水平的抗 A 或抗 B IgG 抗体时,孩子可能有患胎儿和新生儿溶血病 (HDFN) 的风险。对胎儿 ABO 血型进行直接产前测定可以提供有价值的临床信息。目的:在这里,我们报告了一种基于下一代测序 (NGS) 的检测方法,用于预测产前 ABO 血型。材料与方法:从孕12周到35周共检测了26名孕妇的26份血浆样本,其中临床样本20份,测试样本6份。使用 2 个引物组对提取的无细胞 DNA 进行 PCR 扩增,然后进行 NGS。NGS 数据通过 2 种不同的方法(FASTQ 分析和 grep 搜索)进行分析,以确保可靠的结果。将胎儿 ABO 预测与已知的血清学婴儿 ABO 类型进行比较,该类型可用于 19 个样本。结果: 19 个可预测样本中的 19 个具有一致性,其中表型信息可用以及何时通过 2 种方法进行分析。对于已接种疫苗的孕妇(n = 20),预测有 12 个胎儿发生 HDFN 的风险,预测 7 个胎儿没有风险;临床样本的一项结果无法确定。克隆和测序揭示了一种新的变体,它具有与 ABO*O.01.24 相同的单核苷酸变异,并具有额外的 c.220C>T 替代。在 6 个测试样本中发现了一个额外的不确定结果,它是由母体杂合性引起的。由于杂合 ABO 基因或母体杂合性,这 2 个无法确定的样本显示出对测定的限制。结论:我们开创了一种基于 NGS 的胎儿 ABO 预测分析,该分析基于来自母体血浆的无细胞 DNA 分析,并证明了其在少量样本中的应用。基于变异频率和 ABO*O.01/ABO*O.02 杂合子频率的计算,我们估计我们可以在即将到来的 O 型孕妇临床样本中大约 95% 的胎儿 ABO 类型是可靠的。尽管 ABO 血型存在巨大的遗传变异,但许多变异很少见,产前 ABO 预测是可能的,并为预防 ABO HDFN 增加了有价值的早期信息。我们开创了一种基于 NGS 的胎儿 ABO 预测分析,该分析基于来自母体血浆的无细胞 DNA 分析,并在少数样本中证明了其应用。基于变异频率和 ABO*O.01/ABO*O.02 杂合子频率的计算,我们估计我们可以在即将到来的 O 型孕妇临床样本中大约 95% 的胎儿 ABO 类型是可靠的。尽管 ABO 血型存在巨大的遗传变异,但许多变异很少见,产前 ABO 预测是可能的,并为预防 ABO HDFN 增加了有价值的早期信息。我们开创了一种基于 NGS 的胎儿 ABO 预测分析,该分析基于来自母体血浆的无细胞 DNA 分析,并在少数样本中证明了其应用。基于变异频率和 ABO*O.01/ABO*O.02 杂合子频率的计算,我们估计我们可以在即将到来的 O 型孕妇临床样本中大约 95% 的胎儿 ABO 类型是可靠的。尽管 ABO 血型存在巨大的遗传变异,但许多变异很少见,产前 ABO 预测是可能的,并为预防 ABO HDFN 增加了有价值的早期信息。我们估计,在即将到来的 O 型孕妇临床样本中,大约 95% 的胎儿都可以确定可靠的 ABO 类型。尽管 ABO 血型存在巨大的遗传变异,但许多变异很少见,产前 ABO 预测是可能的,并为预防 ABO HDFN 增加了有价值的早期信息。我们估计,在即将到来的 O 型孕妇临床样本中,大约 95% 的胎儿都可以确定可靠的 ABO 类型。尽管 ABO 血型存在巨大的遗传变异,但许多变异很少见,产前 ABO 预测是可能的,并为预防 ABO HDFN 增加了有价值的早期信息。
更新日期:2020-01-01
down
wechat
bug