Dalian Blood Centre, Dalian, China.
Institute of Cytobiology, Dalian Medical University, Dalian, China.
PLoS One. 2022 Apr 1;17(4):e0266332. doi: 10.1371/journal.pone.0266332. eCollection 2022.
To assess the efficacy of Y-chromosome mini-STR-based next-generation sequencing (NGS) for non-invasive prenatal paternity testing (NIPPT).
DNA was extracted from the plasma of 24 pregnant women, and cell-free fetal DNA (cffDNA) haplotyping was performed at 12 Y-chromosome mini-STR loci using the Illumina NextSeq 500 system. The cffDNA haplotype was validated by the paternal haplotype. Subsequentlly, the paternity testing parameters were attributed to each case quantitatively.
The biological relationship between the alleged fathers and infants in all 24 family cases were confirmed by capillary electrophoresis (CE). The Y-chromosome mini-STR haplotypes of all 14 male cffDNA were obtained by NGS without any missing loci. The alleles of cffDNA and paternal genomic DNA were matched in 13 cases, and a mismatched allele was detected at the DYS393 locus in one case and considered as mutation. No allele was detected in the 10 female cffDNA. The combined paternity index (CPI) and probability of paternity calculation was based on 6 loci Y-haplotype distributions of a local population. The probability of paternity was 98.2699-99.8828% for the cases without mutation, and 14.8719% for the case harboring mutation.
Our proof-of-concept study demonstrated that Y-chromosome mini-STR can be used for NGS-based NIPPT with high accuracy in real cases, and is a promising tool for familial searching, paternity exclusion and sex selection in forensic and medical applications.
评估基于 Y 染色体微 STR 的下一代测序（NGS）在非侵入性产前亲子鉴定（NIPPT）中的功效。
从 24 名孕妇的血浆中提取 DNA，并使用 Illumina NextSeq 500 系统在 12 个 Y 染色体微 STR 位点上进行无细胞胎儿 DNA（cffDNA）单体型分析。通过父系单体型验证 cffDNA 单体型。随后，定量地将亲子鉴定参数归因于每个案例。
通过毛细管电泳（CE）确认了所有 24 个家庭案例中被指控父亲与婴儿之间的生物学关系。通过 NGS 获得了所有 14 名男性 cffDNA 的 Y 染色体微 STR 单体型，没有任何缺失的位点。在 13 个案例中，cffDNA 和父系基因组 DNA 的等位基因相匹配，在一个案例中检测到 DYS393 位点的错配等位基因，被认为是突变。在 10 名女性 cffDNA 中未检测到等位基因。基于当地人群的 6 个 Y 单倍型分布，计算了综合亲子鉴定指数（CPI）和亲子鉴定概率。无突变病例的亲子鉴定概率为 98.2699%-99.8828%，突变病例为 14.8719%。
本概念验证研究表明，Y 染色体微 STR 可用于基于 NGS 的 NIPPT，在真实案例中具有高精度，是法医学和医学应用中家族搜索、亲权排除和性别选择的有前途的工具。
