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目的探讨WT1突变致慢性肾功能衰竭产前基因诊断的方法。方法北京大学第一医院于2007年12月对1例先证者家系进行详细的遗传咨询和基因突变分析,提取患儿母亲第2胎孕20周羊水细胞基因组DNA,根据先证者基因突变分析,PCR扩增相应基因相应外显子检测胎儿突变情况,PCR扩增SRY联合核型分析检测胎儿性别,通过3个X染色体微卫星标记(AR、DXS6797和DXS6807)连锁分析除外羊水中母体细胞污染的可能。结果先证者基因突变分析结果为WT1基因IVS9+5G>A杂合突变,同时合并NPHS2第7外显子g.860A>G杂合突变。核型分析显示,患儿核型为46XY。家系突变分析显示,患儿母亲携带NPHS2第7外显子g.860A>G杂合突变,无WT1基因IVS9+5G>A杂合突变,父亲未发现异常。患儿母亲第2胎产前羊水细胞基因组DNA检测显示,胎儿无WT1和NPHS2相应位点突变。PCR扩增SRY基因和核型分析均显示胎儿为女性,连锁分析显示羊水细胞中无母体细胞污染。结论本研究建立了WT1突变所致慢性肾衰竭产前基因诊断的方法。
Objective To investigate the method of prenatal gene diagnosis of chronic renal failure caused by WT1 mutation. Methods The First Hospital of Peking University conducted a detailed genetic counseling and gene mutation analysis on a progenitor pedigree in December 2007 to extract the genomic DNA of amniotic fluid cells of the second gestational age of pregnant women with mothers for the first 20 weeks. According to the gene mutation analysis The genotypes of fetuses were detected by PCR amplification of the corresponding exons. The sex of the fetus was detected by PCR amplification combined with karyotype analysis. The three X chromosome microsatellite markers (AR, DXS6797 and DXS6807) Possible. Results The proband gene mutation analysis showed that the WT1 gene was a heterozygous mutation of IVS9 + 5G> A, and gK60A> G heterozygous mutation of exon 7 of NPHS2. Karyotype analysis showed that children with karyotype 46XY. Family mutation analysis showed that mothers with mothers with NPHS2 exon 7 g.860A> G heterozygous mutation, no WT1 gene IVS9 +5 G> A heterozygous mutation, the father found no abnormalities. The second prenatal amniotic fluid cell genomic DNA test showed that the fetus had no WT1 and NPHS2 mutation. PCR amplification of SRY gene and karyotype analysis showed that the fetus was female and the linkage analysis showed no maternal cell contamination in amniotic fluid cells. Conclusion This study established a method of prenatal gene diagnosis of chronic renal failure induced by WT1 mutation.