多重连接探针扩增及全基因组芯片分析孤独症患者SHANK3及UBE3A等热点基因拷贝数变异初步研究

摘要

目的 研究SHANK3,UBE3A等热点基因拷贝数变异(CNV)与孤独症的相关性.方法 对75名孤独症患儿及112名健康父母和30名正常对照进行研究,利用多重连接探针扩增(MLPA)及全基因组芯片重点对22q13区域(SHANK3基因)、15q11-13 (UBE3A,GABRB3基因)、15q13微缺失区域及CHRNA7基因、16p11微缺失区域等区域进行基因组DNA拷贝数变异检测.结果 ① MLPA分析显示,75名孤独症患儿有6名存在22q13区域的SHANK3基因第15外显子杂合缺失(8.0%,6/75),正常组缺失率为1.4%(2/142),两组间差异有统计学意义(P<0.05);②对6名SHANK3杂合缺失的患儿及6名正常对照进行全基因组芯片分析显示,孤独症患儿CNV变异总数和所涉及的染色体长度都远远高于对照组,在第1,9,15,16,21,22号染色体CNV变化较大.结论 高通量全基因拷贝数变异研究有助于孤独症研究,22q13及SHANK3基因可作为孤独症热点区域重点研究.%Objective To study the correlation of copy number variation in SHANK3,UBE3A and other autism hot gene among children with autism. Metbods 75 children with autism and 112 healthy parents and 30 normal controls were investigated by using multiplex ligation-dependent probe amplification (MLPA) and whole-genomic array method. The CNV of 22q13 region (SHANK3 gene), 15q11-13 (including UBE3A,GABRB3 gene),15q13 microdeletion and 16p11 microdeletion were mainly focused on in this study. Results ① MLPA analysis showed that 6 of 75 children with autism had a significantly higher heterozygous ratio of Exon 15 deletion of SHANK3 gene (6/75,8. 0％) than those normal controls (2/142,1. 4％ ) (P＜0. 05) ;②Whole genome array analysis of 6 autism with heterozygous deletion of SHANK3 and 6 normal contorl showed the total number of CNV variation and length of the CNV change in chromosome 1,9,15,16, 21, 22 in autistic children were far higher than the control samples.Conclusion The high-throughput genomic copy number variation research is helpful to found more information of autism,22q13 region and SHANK3 gene could serve as hot spots in autism research.