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单细胞巢式PCR诊断软骨发育不全 被引量:4

Diagnosing achondroplasia by single cell nested-PCR
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摘要 目的 探讨单细胞水平诊断软骨发育不全 ( achondroplasia,ACH)的可靠性 ,为开展 ACH的胚胎植入前遗传学诊断 ( preimplantation genetic diagnosis,PGD)打下基础。方法 采用巢式 PCR扩增单淋巴细胞及单卵裂球的成纤维细胞生长因子受体 3基因的高发突变位点 G380 R区域 ,用限制酶 Bfm 消化 PCR产物 ,10 %聚丙烯酰胺凝胶电泳检测。结果 单淋巴细胞的 PCR扩增成功率为 90 .4 % ,等位基因脱扣发生率为 8.2 % ,诊断准确率为 91.8% ;单卵裂球的扩增成功率为 75 .4 %。结论 单细胞巢式 PCR诊断 ACH是比较稳定。 Objective: To research on the reliability of diagnosing achondroplasia (ACH) on single cell level and to provide a basis for preimplantation genetic diagnosis (PGD). Methods: The high-frequency mutation region G380R of fibroblast growth factor receptor 3(FGFR3) gene was amplified by nested-PCR with single lymphocyte and single blastomere. The products of PCR were digested by restriction enzyme Bfm I, then the digested products were detected by 10% polyacrylamida gel eledrophoresis (PAGE). Results: The amplification success rate, allele dropout rate and correct diagnosis rate of single lymphocyte's PCR were 90.4%, 8.2% and 91.8%, respectively. The amplification success rate of single blastomere was 75.4%. Conclusion: The diagnosis of ACH by single cell nested-PCR is comparatively stable and reliable.
出处 《中华医学遗传学杂志》 EI CAS CSCD 2003年第3期228-231,共4页 Chinese Journal of Medical Genetics
基金 湖南省社会发展科技项目基金 (1 0 1 3- 8)~~
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