摘要
目的探索唐氏综合征(Down’s Syndrome)的快速诊断方法。方法采用细胞遗传学分析技术对35例唐氏综合征患者(包括4例羊水标本和31例外周血标本)进行核型分析。同时采用荧光定量PCR技术对35例唐氏综合征患者和371例正常对照(含32例羊水标本和339例外周血标本)21号染色体上的D21S11位点进行PCR扩增检测。其中对10例未培养的羊水细胞(4例唐氏综合征患者和6例正常对照)进行荧光原位杂交分析。将唐氏综合征患者的荧光定量PCR检测结果和荧光原位杂交检测结果与细胞遗传学分析结果进行比较。结果35例唐氏综合征患者经细胞核型分析证实,其中嵌合体4例(含1例羊水标本),10例未培养的羊水细胞用荧光原位杂交法检测出唐氏综合征患者5例,经羊水细胞培养,核型分析证实其中4例为唐氏综合征患者。通过对正常对照组研究发现杂合子个体D21S11位点两个等位基因扩增产物的荧光强度的比值接近1,而疾病组12例病人标本,两个相同型别等位基因和第三条等位基因的扩增产物荧光强度的比值在2左右。19例病人标本扩增产物显示第三条D21S11位点等位基因带。4例嵌合体标本,荧光定量PCR检测结果和正常对照一致,无法区分。结论荧光原位杂交技术和荧光定量PCR技术均具备快速,仅需要微量的扩增起始材料的特点,可以用于唐氏综合征的快速产前诊断。但荧光原位杂交技术和荧光定量PCR技术的特异性与敏感性均有待提高。
Objective To explore the rapid diagnosis assay of Down Syndrome. Methods 35 patients with Down Syndrome, including 4 amniofic fluid and 31 peripheral blood samples were analyzed by the cytogenetic methods. At the same time, 35 patients and 371 controls were tested by fluorescent quantitative PCR amplification of D21 S11 STR locu. The control group was composed of normal amniotic fluid samples （ n = 32） and peripheral blood samples （ n = 339）. And 10 uncultured amniotic fluid samples, containing 4 Down Syndrome patients and 6 controls, were detected by fluorescence in situ hybridization. The PCR and hybridization results were compared. Results There were 35 Down Syndrome patients with 4 chimeras verified by the cytogenetic assay. There were 5 out of 10 uncultured amniotic fluid samples were abnormal detected by fluorescence in situ hybridization. The PCR analysis results in controls showed the fluorescent intensity ratios of amplification products of two alleles of heterozygous samples on D21S11 locus were near 1 to 1.12 out of 35 patients showed a ＂diallehc＂ pattern and the influorescent ratio of two alleles were near 2. The PCR product of DNA from 19 patients with Trisomy 21 presented the third allele. Four chimeras couldn＇t be diagnosed by this method. Condusion Since the fluorescent quantitative polymerase chain reaction and fluorescence in situ hybridization methods were rapid and accurate and only small amount of starting material was needed, they could be applied in rapid prenatal diagnosis of Down Syndrome.
出处
《中国实验诊断学》
2006年第5期540-544,共5页
Chinese Journal of Laboratory Diagnosis
关键词
唐氏综合征
核型分析
荧光原位杂交
荧光定量PCR
产前诊断
Down Syndrome
karyotype analysis
fluorescence in situ hybridization
fluorescent quantitative polymerase chain reaction
rapid diagnosis
