7例严重联合免疫缺陷高危儿羊膜腔穿刺产前诊断结果分析.docVIP

7例Wiskott-Aldrich综合征高危儿产前诊断结果分析 摘要 目的：探讨羊水脱落细胞基因分析及脐静脉穿刺脐血WAS蛋白检测在Wiskott-Aldrich综合征（WAS) 高危儿产前诊断中的意义。材料和方法：2008年至2010年我院经WAS蛋白（WASp）流式检测和基因分析明确诊断的7例WAS患儿。详细记录先证者病史，进行家族相关亲属基因检测，建立7个WAS家系图谱，确诊15个异常基因携带者。2008年至2011年间对7个家系中携带异常基因的7个高危孕妇于孕18-20周经羊膜腔穿刺抽取羊水。其中部分羊水提取细胞DNA，经PCR扩增WASp基因。 PCR产物进行双向序列重复测定。其中1例高危孕妇孕28周采集脐带血，采用流式细胞术检测WASp。培养羊水中胎儿脱落细胞, 采用原位制片G带染色技术进行染色体分析。例穿刺均成功，羊水细胞培养成功率％羊amniotic fluid exfoliated cells and WASp detection from cord blood in prenatal daiagnosis of high-risk fetus with Wiskott-Aldrich syndrome. Material and method: Seven patients with Wiskott-Aldrich syndrome were diagnosed by gene analysis and WASP detected by flow cytometry from 2008 to 2010. After detailed inquiry for medical history and gene analysis of related family members, seven pedigree trees were drawn, including 15 carriers of abnormal genes. From 2008 to 2011, seven samples of amniotic cell gotten by amniocentensis were collected from seven high-risk pregnant women with abnormal gene during 18 to 20 gestational weeks. WASP gene was amplified by polymerase chain reaction (PCR) from DNA of amniotic cell gotten and sequencing was performed directly on the PCR products in forward and reversely. Embryo blood sample was collected from one hig-risk fetus by needle puncture of umbilical blood vessel and WASP expression was detected by flow cytometry. Karyotyping was performed in amniotic cell gotten cultivated by orthotope slice and G band staining. Gene analysis of WASP、WASP expression detected by flow cytometry and evaluation of immune function were reexamined in high-risk fetus after delivery. Result: Amniocentensis and culture of amniotic cell gotten all successed in seven fetus. Gene analysis and karyotyping showed that one male fetus and four female fetus were normal and two female fetus were carriers. WASP expression detected from embryo blood sample of the patient showed normal. After delivery, the result of gene analysis，WASP detection and evaluation of immune function was same wi