个体化用药基因检测-十堰太和医院.ppt

Human Genome Project research showed that 99.9 percent of individual genes is the same, but there are sequences in a very small (0.1 percent) of genetic differences, which mainly were SNPs. SNPs (single nucleotide polymorphism) most commonly refer to single-base differences in DNA among individuals. Polymorphisms are usually defined as sites where the less allele has a frequency of at least 1% in the population. SNPs present in the entire human genome, may be each 100-300bp existed a SNP, estimated that the total number in the millions, but for some purposes rarer variants are important as well. Some useful online SNP databases /genesnps/ /snplist.cfm /entrez/query.fcgi?db=PubMed 检测基因多态性的实验方法 PCR-SSCP PCR-RLFP PCR-SSO PCR-ASP Direct sequencing SNaPshot dHPLC Real-time PCR PCR-RFLP Amplify the DNA fragment by PCR, then using specific enzyme digestion to cut into different fragments, gel-electrophoresis to distinguish. Different alleles of the restriction site distribution, have a length of DNA fragments of different bands. ---CACAGGAAGCCTGCAGTCCTGGAA--- Pst I ---CACAGGAAGCCTGTAGTCCTGGAA--- TaqMan原理 在PCR扩增反应时，一对两端有不同荧光标记的特异探针来识别不同的等位基因（allele1和allele2），5’端为报告荧光基团，3’端为淬灭荧光基团。 当探针以完整形式存在时，由于能量共振转移，荧光基团只发出微弱荧光。特异的探针与相应的等位基因杂合后，DNA聚合酶发挥5’到3’外切酶活性，将报告荧光基团切割下来，脱离了3’端淬灭荧光基团的淬灭作用，从而发出荧光。 SCI论文的框架结构 多态性meta分析论文的模式 类似八股文的模拟 专业知识的填充 统计学知识的运用 SNPmeta SNPmeta SNPmeta SNPmeta SNPmeta 1057179657@QQ.COM 谢谢！ 流行病学研究方法 SNPmeta 基因多态性与疾病的关联研究，通常采用病例-对照研究的方法，检测SNP的状态在病例组和对照组之间是否有差异。经过统计学处理，得到Odds Ratio(ORs)等信息。 SNPmeta 效应指标的选择 基因模型的选择 假设基因突变为A到B，则三种基因型分别为AA，AB，BB。 假定AA为野生纯合基因型，则AB为突变杂合基因型，BB为突变纯合基因型。 则常用的基因模型为以下5种： BB vs. AA (homozygote) AB vs. AA (heterozygote) BB + AB vs. AA (dominant model) BB vs. AB + AA (recessive model) Allele B vs. Allele A SNPmeta 被dbSNP数据库收录的每一个SNP均会被分配给一个不重复的，以rs加上阿拉伯数字组成的号码，如rs1799793。 检索SNP常用的网站是： dbSNP数据库：/projects/SNP SNPmeta SNP信息的查询 /projects/SNP SNPmeta SNP信息的查询 制作（基因多态性）Meta分析的基本步骤1、提出问题，制定研究计划2、文献检索