应用变性高效液相色谱技术检测三个遗传性多发性外生骨疣家系的EXT1与EXT2基因突变  

EXT1 and EXT2 mutation identified by denaturing high performance liquid chromatograph in three families with hereditary multiple exostoses

作　　者：张萌 刘世国 李飞峰 周万灏 金孝华 马旭 

机构地区：[1]国家人口计生委科研所遗传室,北京100081

出　　处：《中华医学遗传学杂志》2007年第6期646-651,共6页Chinese Journal of Medical Genetics

摘　　要：目的建立一种应用变性高效液相色谱（denaturing high performance liquid chromatograph,DHPLC）技术检测遗传性多发性外生骨疣（hereditary multiple exostosis,EXT）基因突变的新方法,并研究3个EXT家系的基因突变情况。方法扩增EXT致病基因的所有外显子区及部分内含子与外显子交界区,联合连锁分析、DHPLC筛查及测序的方法进行分析。结果3个EXT家系中,共检测到两处已知EXT2基因的剪接位点突变IVS2＋1G〉A、IVS7＋1G〉T,一处28C〉A变异和一处EXT1基因的IVS4＋66G〉A变异。结论EXT2基因的2个剪接位点突变分别是引起相应EXT家系的致病原因,应用DHPLC技术检测遗传性疾病基因变突是一种自动化、高通量、灵敏、快速且简便经济的好方法。Objective To develop a new denaturing high performance liquid chromatograph （DHPLC）-based method to screen patients with EXT gene mutation and to study the gene mutation in three families with multiple exostoscs. Methods All the exons of EXT gene, including the intro-exon boundaries, were amplified by PCR. Linkage analysis and DHPLC screening were carried out to identify the mutations. DNA sequencing was used to confirm the muta- tions. Results Two known splice site mutations, IVS2 ＋ 1 G 〉 A and IVS7 ＋ 1 G 〉 T, and two SNPs have been detected in EXT2 or EXT1 gene. Conclusion The transversions of IVS2 ＋ 1 G 〉 A and IVS7 ＋ 1 G 〉 T in EXT2 gene are suggested to be the disease-causing mutations and the DHPLC is a high throughout, sensitive, simple, quick, economical method to screen gene mutation in hereditary multiple exostosis.

关 键 词：变性高效液相色谱 遗传性多发性外生骨疣 EXT基因 突变分析 

分 类 号：R686[医药卫生—骨科学]