拷贝数变异（CNV）及研究策略和技术.pdfVIP

医学遗传学新方向 拷贝数变异（CNV ）的研究策略和技术 王 颖 市场部经理 上海天昊生物科技有限公司 Copy-number variant (CNV)：A segment of DNA that is 1 kb or larger and is present at a variable copy number in comparison with a reference genomeinsertions, deletions and duplications. Copy-number polymorphism (CNP)：A CNV that occurs in more than 1% of the population. CNV与人类疾病密切相关 CNV与人类疾病相关的部分报道 ※Mendelian Disorders: Dup(7)(q11.23) syndrome: duplication in 7q11.23 Williams-Beuren syndrome: deletions in 7q11.23 22q11 deletion syndrome: 22q11.21-22q11.23 deletion Charcot-Marie Tooth neuropathy Type 1A: duplications of peripheral myelin protein- 22 at chromosome region 17p11.2 ※Complex Disease: Systemic lupus erythematosus (SLE): Complement component 4 (C4) and Fcγ Receptor 3B (FCGR3B) HIV/AIDS susceptibility: CCL3L1; Rheumatoid Arthritis: CCL3L1; Colonic Crohn Disease: DEFB4 Autism: 16p11.2 De novo deletion (OR=100); 16p11.2 De novo duplication(OR=16) Schizophrenia: 1q21.1 De novo deletion(OR=15);15q13.3 De novo deletion (OR=12) ※Drug Response: CYP2D6, SULT1A1 基因剂量效应与疾病 儿童先天性智力残疾和/或发育迟缓的CNV研究 在人类基因组中检测结构变异 Agilent Affymetrix Illumina Feuk L et al, Nat Rev Genet. 2006 7:85-97 DGV的CNV信息 现有目标基因拷贝数检测方法: 1、Real-Time qPCR 2、FISH: Fluorescent In Situ Hybridization 3、MLPA：Multiplex ligation-dependent probe amplification 4 、LOH analysis: Microsatellite Genotyping/SNP Genotyping 5、Analysis of Abnormal Transmission of Genetic Markers 6、Southern Blotting 7、MAPH: multiplex amplifiable probe hybridization 8、QMPSF：Quantitative multiplex PCR of short fluorescent fragments 现有技术的不足之处 1、Real-Time qPCR: 通量低，稳定性及准确性不高 2、FISH: 成本高，通量小，时间长，对