Investigation of single nucleotide polymorphism loci susceptible to degradation by ultraviolet ligh
Investigation of single nucleotide polymorphism loci susceptible to degradation by ultraviolet ligh
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易受紫外线降解的单核苷酸多态性位点的研究
DOI：
10.1016/j.jflm.2016.08.002
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发表时间：
2016
影响因子：
1.5
通讯作者：
Kibayashi K
中科院分区：
文献类型：
--
作者：
Machida M;Taki T;Shimada R;Kibayashi K
生物体液中的DNA经常被环境因素降解。鉴于单核苷酸多态性（SNP）分析需要比短串联重复序列（STR）分析更短的扩增子，它们在使用降解样品进行人类鉴定中的应用最近引起了人们的关注。尽管各种SNP位点被用于分析降解样品，但尚不清楚哪种更合适。为了表征和鉴定对降解敏感或抗性的SNP位点，我们通过暴露于紫外线（UV）不同持续时间（254 nm，5、15、30、60或120 min）来人工降解从11名志愿者的口腔拭子获得的DNA，并分析所得SNP位点。使用凝胶电泳、STR和SNP分析评估DNA降解。DNA片段发生在5分钟内的紫外线照射，成功的STR和SNP分析随着持续时间的增加而减少。然而，73%的SNP位点仍然正确地检测到DNA样品照射120分钟，剂量，使STR位点检测不到。不成功的SNP分型和邻近SNP的核苷酸的碱基调用失败被追溯到rs1031825，我们发现该SNP对紫外光敏感。当比较STR和SNP基因座的检测效率时，SNP分型比STR分型更成功，这使得使用降解DNA时更有效。然而，在解释降解DNA的SNP分析时，谨慎使用rs1031825是很重要的。
DNA in biological fluids is often degraded by environmental factors. Given that single nucleotide polymorphism (SNP) analyses require shorter amplicons than short tandem repeat (STR) analyses do, their use in human identification using degraded samples has recently attracted attention. Although various SNP loci are used to analyze degraded samples, it is unclear which ones are more appropriate. To characterize and identify SNP loci that are susceptible or resistant to degradation, we artificially degraded DNA, obtained from buccal swabs from 11 volunteers, by exposure to ultraviolet (UV) light for different durations (254 nm for 5, 15, 30, 60, or 120 min) and analyzed the resulting SNP loci. DNA degradation was assessed using gel electrophoresis, STR, and SNP profiling. DNA fragmentation occurred within 5 min of UV irradiation, and successful STR and SNP profiling decreased with increasing duration. However, 73% of SNP loci were still detected correctly in DNA samples irradiated for 120 min, a dose that rendered STR loci undetectable. The unsuccessful SNP typing and the base call failure of nucleotides neighboring the SNPs were traced to rs1031825, and we found that this SNP was susceptible to UV light. When comparing the detection efficiencies of STR and SNP loci, SNP typing was more successful than STR typing, making it effective when using degraded DNA. However, it is important to use rs1031825 with caution when interpreting SNP analyses of degraded DNA.