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目的 建立一个快速、准确诊断 2 1 三体综合征 (2 1 三体 )方法。方法 取 2 6例 2 1 三体患儿及 2 0例正常人DNA标本 ,引用一对引物同时PCR扩增两个同源基因片断 :2 1号染色体长臂的人肝型磷酸果糖激酶基因 (PFKL CH2 1)及 1号染色体的人肌型磷酸果糖激酶基因 (PFKM CH1) ,用SYBRGreenI荧光染色 ,琼脂糖凝胶电泳、凝胶成像系统分析软件测光密度进行定量分析。结果 2 4个PCR循环检测两个同源基因扩增产物光密度之比 :病例组为 1.6 1± 0 .18;正常人组为 1.0 1± 0 .0 6 ,两组比值分布无重叠区 ,诊断结果与染色体核型分析一致 ,约 4h可完成。结论 SYBRGreenI荧光同源基因定量PCR方法省时、简单、快捷、准确诊断 2 1 三体 ,为临床及产前诊断 2 1 三体提供一种新的诊断方法。
Objective To establish a rapid and accurate method for the diagnosis of trisomy 21 (21 trisomy). Methods Twenty-six children with 21 trisomy and 20 normal controls were enrolled in this study. A pair of primers was used to simultaneously amplify two homologous gene fragments: the human liver-type phosphofructokinase gene PFKL CH2 1) and human chromosome-type phosphofructokinase gene (PFKM CH1) on chromosome 1 were quantitatively analyzed by SYBR Green I fluorescent staining, agarose gel electrophoresis and gel imaging system software. Results 24 PCR cycles were performed to detect the ratio of optical density of the two amplified products: 1.6 1 ± 0.18 in the case group and 1.0 1 ± 0.06 in the normal group, with no overlap in the distribution of the two groups, Diagnostic results and chromosome karyotype analysis, about 4h can be completed. Conclusion SYBRGreenI fluorescent homologous gene quantitative PCR method is a simple, rapid and accurate method for the diagnosis of 21 trisomy. It provides a new diagnostic method for clinical and prenatal diagnosis of 21 trisomy.