常用STR基因座在人消化系统肿瘤组织中的变异分析
发布时间：2019-05-22 05:51
【摘要】： 目的:对常用STR基因座在人消化系统肿瘤组织中的变异情况进行系统研究和分析,对变异的类型、规律等进行总结,筛选出变异率小、稳定性强的STR基因座,并进一步建立验证突变的技术手段。 方法:收集新鲜人消化系统肿瘤组织和对应正常组织99例,用Chelex-100法提取基因组DNA;采用Identifiler试剂盒对所有样本扩增,扩增产物通过ABI3100-Avant型遗传分析仪进行毛细管电泳,收集电泳信息,用Genemapper 3.2检测各位点的基因型,对分型结果进行比较,分别筛选出不同类型的变异样本;根据STR基因座的基因组学和遗传学特征,设计引物,建立用于验证STR变异的技术手段。 结果:对于99例所收集样本,均成功提取了基因组DNA,经STR分型试剂盒对所有样本扩增,获得的扩增产物进行毛细管电泳检测,结果表明有49例样本发生了变异,总变异率近50%。变异的类型包括增加等位基因、基因型变更、完全杂合性丢失和部分杂合性丢失,其中增加等位基因37例,基因型变更3例,完全杂合性缺失14例,部分杂合性丢失83例。 结论:四种变异类型在15个STR基因座中都有分布,其中变异频率最大的STR基因座依次为FGA、D18S51、D5S818、D13S317和D8S1179。TH01基因座、D2S1338基因座、TPOX基因座变异频率最低,具有较好的稳定性。所建立的PCR体系可以用于验证肿瘤组织中STR基因座是否发生了突变。
[Abstract]:Objective: to systematically study and analyze the variation of STR loci in human digestive system tumors, summarize the types and laws of variation, and screen out the STR loci with low mutation rate and strong stability. And further establish the technical means to verify the mutation. Methods: 99 cases of fresh human digestive system tumor tissues and corresponding normal tissues were collected and genomic DNA; was extracted by Chelex- 100 method. All samples were amplified by Identifiler kit, and the amplified products were detected by capillary electrophoresis with ABI3100- Avant genetic analyzer. The electrophoresis information was collected, the genotypes of each point were detected by Genemapper 3.2, and the typing results were compared. Different types of variation samples were screened out. According to the genomics and genetic characteristics of STR loci, primers were designed to establish a technical means to verify STR variation. Results: for 99 samples collected, genomic DNA, was successfully extracted and all samples were amplified by STR typing kit. The amplified products were detected by capillary electrophoresis. The results showed that 49 samples had mutated. The total variation rate is nearly 50%. The types of variation included increased alleles, genotypic changes, complete loss of heterozygosity and loss of partial heterozygosity, including 37 cases of increased alleles, 3 cases of genotypic changes, 14 cases of complete loss of heterozygosity and 83 cases of partial loss of heterozygosity. Conclusion: the four variation types are distributed in 15 STR loci, among which the STR loci with the largest frequency of variation are FGA,D18S51,D5S818,D13S317 and D8S1179.TH01 loci, D2S1338 loci and TPOX loci have the lowest variation frequency. It has good stability. The established PCR system can be used to verify the mutation of STR locus in tumor tissues.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：D919
本文编号：2482729
[Abstract]:Objective: to systematically study and analyze the variation of STR loci in human digestive system tumors, summarize the types and laws of variation, and screen out the STR loci with low mutation rate and strong stability. And further establish the technical means to verify the mutation. Methods: 99 cases of fresh human digestive system tumor tissues and corresponding normal tissues were collected and genomic DNA; was extracted by Chelex- 100 method. All samples were amplified by Identifiler kit, and the amplified products were detected by capillary electrophoresis with ABI3100- Avant genetic analyzer. The electrophoresis information was collected, the genotypes of each point were detected by Genemapper 3.2, and the typing results were compared. Different types of variation samples were screened out. According to the genomics and genetic characteristics of STR loci, primers were designed to establish a technical means to verify STR variation. Results: for 99 samples collected, genomic DNA, was successfully extracted and all samples were amplified by STR typing kit. The amplified products were detected by capillary electrophoresis. The results showed that 49 samples had mutated. The total variation rate is nearly 50%. The types of variation included increased alleles, genotypic changes, complete loss of heterozygosity and loss of partial heterozygosity, including 37 cases of increased alleles, 3 cases of genotypic changes, 14 cases of complete loss of heterozygosity and 83 cases of partial loss of heterozygosity. Conclusion: the four variation types are distributed in 15 STR loci, among which the STR loci with the largest frequency of variation are FGA,D18S51,D5S818,D13S317 and D8S1179.TH01 loci, D2S1338 loci and TPOX loci have the lowest variation frequency. It has good stability. The established PCR system can be used to verify the mutation of STR locus in tumor tissues.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：D919
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相关期刊论文 前2条
1 刘开会,季安全,邱宁,王坚,王传海,郭燕霞,常彩琴,王占海,王建国,曹侠;肿瘤组织9个STR基因座及Amelogenin基因座突变[J];中国法医学杂志;2002年02期
2 方建新;李成涛;肖立;;13个STR位点在人消化系统肿瘤组织中的变异分析[J];法医学杂志;2007年04期
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