CAJ | 학술논문

目的探讨Wiskott-Aldrich综合征(WAS)高危孕妇羊水脱落细胞基因分析及脐静脉穿刺脐血WAS蛋白(WASP)检测在WAS高危儿产前诊断中的意义.方法 2008年至2010年我院经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.2008年至2011年对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WAS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,采用原位制片及G带染色技术进行染色体核型分析.产后采集高危儿外周血进行基因分析和WASP检测验证产前诊断结果.结果 7例WAS高危儿羊膜腔穿刺均成功,羊水细胞培养成功率100％.WAS基因和染色体核型分析结果显示1例正常男性胎儿,4例正常女性胎儿,2例为女性异常基因携带者.1例脐带血流式细胞术检测WASP显示正常表达.7例高危儿均顺利出生,产后WASP和基因分析结果均正常,与产前结果相同.结论羊水脱落细胞核型、基因分析与脐带血WASP检测可为WAS高危孕妇提供可靠的产前诊断服务.
목적 탐토Wiskott-Aldrich종합정(WAS)고위잉부양수탈락세포기인분석급제정맥천자제혈WAS단백(WASP)검측재WAS고위인산전진단중적의의.방법 2008년지2010년아원경WASP류식검측화기인분석명학진단적7례WAS환인.상세기록선증자병사,진행가족상관친속기인검측,건립7개WAS가계도보.2008년지2011년대7개가계중휴대이상기인적7개고위잉부우잉18 ～ 20주경양막강천자추취양수.기중부분양수제취세포DNA,경PCR확증WAS기인.PCR산물진행쌍향서렬중복측정.기중1례고위잉부잉28주채집제대혈,채용류식세포술검측WASP.배양양수중태인탈락세포,채용원위제편급G대염색기술진행염색체핵형분석.산후채집고위인외주혈진행기인분석화WASP검측험증산전진단결과.결과 7례WAS고위인양막강천자균성공,양수세포배양성공솔100％.WAS기인화염색체핵형분석결과현시1례정상남성태인,4례정상녀성태인,2례위녀성이상기인휴대자.1례제대혈류식세포술검측WASP현시정상표체.7례고위인균순리출생,산후WASP화기인분석결과균정상,여산전결과상동.결론 양수탈락세포핵형、기인분석여제대혈WASP검측가위WAS고위잉부제공가고적산전진단복무.
Objective To investigate the value of gene analysis of amniotic fluid exfoliated cells and WASP detection from cord blood in prenatal diagnosis of high-risk fetus with Wiskott-Aldrich syndrome.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 amniocentesis 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 forward and reversely.Embryo blood sample was collected from one high-risk fetus by needle puncture of bilical 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 succeeded in all the seven fetuses.Gene analysis and karyotyping showed that one male fetus and four female fetusse were normal and two female fetuses were carriers. WASP expression detected from embryo blood sample of the patient was normal. After delivery,the result of gene analysis,WASP detection and evaluation of immune function was the same as that of prenatal diagnosis.Conclusion Karyotyping,gene analysis and WASP detection of cord blood can provide reliable service of prenatal diagnosis for high-risk pregnant women with Wiskott-Aldrich syndrome.