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血友病A是人类缺少凝血因子Ⅷ所致的最常见遗传性出血疾病,通过Sou thern blotting和限制片段长度多态性(RFLP)进行携带者检出和产前诊断,已大大改善了对这一疾病的预防。本篇报道应用Ⅷ因子基因的19内含子HindⅡ多态性位点,采用PCR技术作血友病A的产前诊断,使产前诊断时间从2周减至1～2天。取10mg妊娠11周的绒毛膜用分离白细胞中的DNA的方法进行DNA提取,清洗后的绒毛膜样品用蛋白酶K和RNase消化,然后进行酚抽提和沉淀,在25μl的体积中进行靶片段的放大,其中含有0.5μg基因组DNA,1.5mM四种单核苷酸,50pmol寡核苷酸引物,反应体系的缓冲液为16.6mM
Hemophilia A is the most common hereditary bleeding disorder caused by the absence of factor VIII in humans and carrier detection and prenatal diagnosis by Sou thern blotting and restriction fragment length polymorphism (RFLP) have greatly improved the Prevention of a disease. This report uses Ⅷ factor gene 19 intron Hind Ⅱ polymorphic sites, PCR technology for hemophilia A prenatal diagnosis, prenatal diagnosis of time from 2 weeks to 1 to 2 days. 10 mg of chorion for 11 weeks of gestation was used for DNA extraction from DNA isolated from leukocytes. The washed chorionic sample was digested with proteinase K and RNase, followed by phenol extraction and precipitation. The target fragment Containing 0.5 μg of genomic DNA, 1.5 mM of four mononucleotides, and 50 pmol of oligonucleotide primer. The reaction buffer was 16.6 mM