微乳液多重PCR基因芯片法在早孕期无创性胎儿性别诊断中应用.docVIP

微乳液多重PCR基因芯片法在早孕期无创性胎儿性别诊断中应用[摘要]目的探讨微乳液多重PCR基因芯片法在早孕期无创性胎儿性别诊断中的应用价值。方法收集138例4~12孕周孕妇的外周血液样品，提取血浆DNA。选取胎儿DNA Y染色体上的6个特异性序列SRY1、SRY2、SRY3、DYS1、DYS14和DYZ3基因作为男性胎儿标志物，采用微乳液PCR进行扩增，同时标记荧光，以β-globin基因作为阳性内对照。扩增产物与固定在基因芯片上的探针分子杂交，采用ScanArray 5000扫描仪扫描芯片。检测结果与胎儿出生性别进行比较。结果最早的检出样品来自孕31d的孕妇，在78份男性胎儿孕育者血样中，有76份样本6个Y染色体特异序列均为阳性；60份女性胎儿孕育者血样中没有一个Y染色体特异序列阳性。微乳液多重PCR基因芯片法对男性胎儿早孕期检测的灵敏度达到97.4%，特异度达到100%。结论微乳液多重PCR基因芯片法检测孕妇外周血中男性胎儿DNA的方法具有较高的灵敏度和特异度，能够早期鉴定胎儿性别，对性连锁遗传病的筛选具有潜在的应用价值。? [关键词] 胎儿DNA；基因芯片；微乳液；多重PCR；无创性产前诊断? [中图分类号] R714.52 [文献标识码] A [文章编号] 1671-7562(2007)02-0083-05?? Non-invasive determination of fetal gender using microarray ?coupled with microemulsions PCR in first trimester? YAN Ling1,GE Qin-yu2，JIANG Xiao-qing3，JIANG Li?1 （1.School of Clinical Medical Science,Zhongda Hospital，Southeast University，Nanjing 210009,China;2.State Key Laboratory ?of Bioelectronics，Southeast University，Nanjing 210096,China;3.Department of Obstetrics and Gynecology，?Jiangsu Province Hospital of Traditional Chinese Medicine，Nanjing 210029,China）?? Abstract：ObjectiveTo evaluate the method of microarray coupled with microemulsions polymerase chain reaction(PCR)in non-invasive determination of fetal gender in first trimester.MethodsPlasma samples were obtained from 138 pregnant women at 4-12 weeks of gestation.Fetal DNA extracted from the plasma of pregnant women.Then six Y chromosome special sequences SRY1，SRY2，SRY3,DYS1，DYS14 and DYZ3 as the marker of fetal DNA were amplified by multiplex-PCR in microemulsions，and fluorescence was labeled at the same times,β- globin gene as an internal control.The labeled target DNA was hybridized and detected by the capturing DNA probes on a microarray slide，signals were scanned by ScanArray 5000 and analyzed.The results were compared to fetal gender at birth.ResultsThe earliest plasma sample which we could detect in this study was collected on the 31st day after pregnancy.The six Y chromosome sequences were detected in 76 of 78 male c