High resolution size analysis of fetal DNA in the urine of pregnant women by paired-end massively parallel sequencing.
High resolution size analysis of fetal DNA in the urine of pregnant women by paired-end massively parallel sequencing.
复制标题
DOI：
10.1371/journal.pone.0048319
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Lo YM
中科院分区：
文献类型：
--
作者：
Tsui NB;Jiang P;Chow KC;Su X;Leung TY;Sun H;Chan KC;Chiu RW;Lo YM
孕妇尿液中的胎儿DNA，如果存在，将是非侵入性产前诊断胎儿遗传物质的宝贵来源。然而，孕妇尿液中是否存在胎儿DNA一直存在争议。这个问题是由于缺乏适当的技术来检测母体尿液中可能高度降解的胎儿DNA。我们使用大量平行配对端测序来研究母体尿液中的无细胞DNA分子。在妊娠晚期收集了7名孕妇的导尿样本。我们通过鉴定含有胎儿特异性单核苷酸多态性等位基因的测序片段来检测胎儿DNA。单个尿液DNA片段的大小从配对reads的比对位置推断出来。我们测量了胎儿DNA浓度的分数以及母体尿液中胎儿和母体DNA的大小分布。7份产妇尿液中有5份检测到无细胞胎儿DNA，胎儿DNA分数浓度范围为1.92% ~ 4.73%。分娩后，孕妇尿液中检测不到胎儿DNA。与血浆DNA相比，尿细胞游离DNA分子的完整性较差。尿胎DNA片段非常短，最优势的胎儿序列长度在29 ~ 45 bp之间。通过大规模平行测序，我们证实了母体尿液中存在经肾胎儿DNA，并表明尿中胎儿DNA被严重降解。
Fetal DNA in maternal urine, if present, would be a valuable source of fetal genetic material for noninvasive prenatal diagnosis. However, the existence of fetal DNA in maternal urine has remained controversial. The issue is due to the lack of appropriate technology to robustly detect the potentially highly degraded fetal DNA in maternal urine. We have used massively parallel paired-end sequencing to investigate cell-free DNA molecules in maternal urine. Catheterized urine samples were collected from seven pregnant women during the third trimester of pregnancies. We detected fetal DNA by identifying sequenced reads that contained fetal-specific alleles of the single nucleotide polymorphisms. The sizes of individual urinary DNA fragments were deduced from the alignment positions of the paired reads. We measured the fractional fetal DNA concentration as well as the size distributions of fetal and maternal DNA in maternal urine. Cell-free fetal DNA was detected in five of the seven maternal urine samples, with the fractional fetal DNA concentrations ranged from 1.92% to 4.73%. Fetal DNA became undetectable in maternal urine after delivery. The total urinary cell-free DNA molecules were less intact when compared with plasma DNA. Urinary fetal DNA fragments were very short, and the most dominant fetal sequences were between 29 bp and 45 bp in length. With the use of massively parallel sequencing, we have confirmed the existence of transrenal fetal DNA in maternal urine, and have shown that urinary fetal DNA was heavily degraded.
登录
查看更多内容
影响因子：
9.3
作者：
Shekhtman, Eugene M.;Anne, Kalpana;Umansky, Samuil R.
通讯作者：
Umansky, Samuil R.
影响因子：
7.2
作者：
Bianchi, Diana W.;Platt, Lawrence D.;Rava, Richard P.
通讯作者：
Rava, Richard P.
影响因子：
11.5
作者：
Chan, K. C. Allen;Leung, Sing F.;Lo, Y. M. Dennis
通讯作者：
Lo, Y. M. Dennis
影响因子：
3
作者：
Illanes, S.;Denbow, M. L.;Finning, K.
通讯作者：
Finning, K.
影响因子：
3
作者：
Koide, K;Sekizawa, A;Okai, T
通讯作者：
Okai, T