Microarray analysis of cell-free fetal DNA in amniotic fluid: A prenatal molecular karyotype
Microarray analysis of cell-free fetal DNA in amniotic fluid: A prenatal molecular karyotype
复制标题
DOI：
10.1086/423288
复制
发表时间：
2004-09-01
影响因子：
9.8
通讯作者：
Bianchi, DW
中科院分区：
文献类型：
--
作者：
Larrabee, PB;Johnson, KL;Bianchi, DW
通过胎盘穿刺或绒毛取样获得的胎儿细胞的中期核型分析是产前细胞遗传学诊断的当前标准，特别是用于检测21三体。我们以前证明了大量的无细胞胎儿DNA（cffDNA）很容易从羊水（AF）中提取。在这项研究中，我们通过测试AF cffDNA与DNA微阵列杂交以进行比较基因组杂交（CGH）分析的能力来探索AF cffDNA的潜在临床应用。与女性参考DNA相比，从11个男性胎儿中分离的cffDNA显示SRY上的杂交信号显著增加，X染色体标记上的信号降低。从六个雌性胎儿分离的cffDNA与雄性参考DNA相比显示相反。与整倍体女性参考DNA相比，来自三个具有21三体的胎儿的cffDNA在大多数21号染色体标记上具有增加的杂交信号，并且来自具有X单体（特纳综合征）的胎儿的cffDNA在大多数X染色体标记上具有减少的杂交信号。这些结果表明，可以使用CGH微阵列分析从AF提取的cffDNA，以正确鉴定胎儿性别和非整倍性。该技术有助于快速筛查样品的全染色体变化，并可通过提供额外的分子信息来增强标准核型分析技术。
Metaphase karyotype analysis of fetal cells obtained by amniocentesis or chorionic villus sampling is the current standard for prenatal cytogenetic diagnosis, particularly for the detection of trisomy 21. We previously demonstrated that large quantities of cell-free fetal DNA (cffDNA) are easily extracted from amniotic fluid (AF). In this study, we explored potential clinical applications of AF cffDNA by testing its ability to hybridize to DNA microarrays for comparative genomic hybridization (CGH) analysis. cffDNA isolated from 11 male fetuses showed significantly increased hybridization signals on SRY and decreased signals on X-chromosome markers, compared with female reference DNA. cffDNA isolated from six female fetuses showed the reverse when compared with male reference DNA. cffDNA from three fetuses with trisomy 21 had increased hybridization signals on the majority of the chromosome 21 markers, and cffDNA from a fetus with monosomy X (Turner syndrome) had decreased hybridization signals on most X-chromosome markers, compared with euploid female reference DNA. These results indicate that cffDNA extracted from AF can be analyzed using CGH microarrays to correctly identify fetal sex and aneuploidy. This technology facilitates rapid screening of samples for whole-chromosome changes and may augment standard karyotyping techniques by providing additional molecular information.