先天性脊柱骨骺发育不良的产前快速分子诊断  

Rapid molecular prenatal diagnosis of spondyleopiphyseal dysplasia congenita by nested SSP-PCR assay

作　　者：崔英霞[1] 夏欣一[1] 杨滨[1] 卢洪涌[1] 史轶超[1] 梁泉[1] 姚兵[1] 李晓军[1] 黄宇烽[1] 

机构地区：[1]南京军区南京总医院解放军临床检验医学研究所,江苏南京210002

出　　处：《医学研究生学报》2008年第10期1049-1052,共4页Journal of Medical Postgraduates

基　　金：江苏省135工程重点学科基金资助项目(批准号:[2001]34);南京军区南京总医院科研基金资助项目(批准号:2006077)

摘　　要：目的:对先天性脊柱骨骺发育不良提供1种快速分子产前诊断技术。方法:对该患者于孕14周进行羊膜囊穿刺,获取羊水20 ml,提取羊水细胞基因组DNA。选用3个短串连重复(STR)序列位点,进行荧光PCR检测,以排除母源细胞污染。在此基础上,对COL2A1基因的第23外显子进行扩增,并以此为模板,针对COL2A1基因的第23外显子1510G→A的突变进行SSP-PCR。根据是否存在特异性的扩增条带进行判断,对初次扩增的产物测序,以证实巢式SSP-PCR的结果。结果:对D13S 317、D18S51和D21S11位点的STR检测结果排除母源细胞的污染。羊水样本仅在野生型反应管中有扩增产物,表明胎儿是COL2A1基因的第23外显子1510 G的纯合子,未带COL2A1基因的第23外显子1510G→A的突变。测序结果与SSP-PCR结果一致。结论:SSP-PCR技术快速简便、敏感特异,可作为该突变的快速分子诊断。其他已知点突变遗传病的快速分子诊断也可以考虑使用SSP-PCR技术进行。Objective： To report the first rapid molecular prenatal diagnosis of congenital spondyloepimetaphyseal dysplasia（SEDC） performed in China by nested SSP-PCR analysis.Methods： A quick molecular prenatal diagnosis of SEDC was performed by nested SSP-PCR assay on an amniocyte sample collected at 14 weeks of pregnancy.Fetal DNA was first examined by short tandem repeats（STR） to exclude the possible contamination of maternal origin.Subsequently,PCR amplification of exon 23 in the COL2A1 gene was carried out and 1∶1 000 dilution of the primary PCR product used as the template with a protocol of SSP-PCR.Simultaneously,the primary PCR product was sequenced using ABI Prism 3 700 automated sequencer to identify the result of SSP-PCR.Results： Analyses of the polymorphic loci of D21S11,D13S 317 and D18S51 excluded the contamination of maternal origin.SSP-PCR assay revealed the fetus without the 1510G→A transition in exon 23 of the COL2A1 gene,which was confirmed by sequencing directly.Conclusion： The nested SSP-PCR method can be applied to rapid molecular prenatal diagnosis of this mutation,as well as other genetic disorders once a point mutation is known.

关 键 词：先天性脊柱骨骺发育不良 快速分子诊断 产前 COL2A1基因 巢式序列特异引物扩增 序列分析 

分 类 号：R714.55[医药卫生—妇产科学]