摘要
目的基于二代测序平台评估法医常见体液样本的编码区单核苷酸多态性(coding region single nucleotide polymorphism,cSNP)在DNA和RNA水平分型一致性。方法经大量文献检索,选取外周血、精液和阴道分泌液中的8个体液特异性mRNA分子上的25个c SNP位点,基于MiSeq FGx测序平台,分别构建DNA水平的c SNP位点复合检验体系和RNA水平的cSNP位点复合检验体系,共使用45份体液样本(包括14份外周血,15份精液,16份阴道分泌液)进行测序并分析各cSNP位点的分型结果,利用Sanger测序技术对在DNA和RNA水平分型结果不一致的位点进行复核。结果外周血特异的标记在DNA和RNA水平的cSNP分型结果完全一致。在15个精液样本中,精液特异标记TGM4基因上rs1995640、rs1995641均有3例分型不一致。在16个阴道分泌液样本中,阴道分泌液特异性标记MUC22基因上rs3869098、rs10947121、rs12110470分别出现2例、1例、2例检验结果不一致的情况。结论本研究基于MiSeq FGx测序平台结合Sanger测序技术检验的25个具有体液组织特异性的cSNP位点中,共有20个位点在DNA和RNA水平分型结果一致,TGM4和MUC22基因上共5个位点在相应体液个别样本的DNA和RNA水平上有差异,本研究为cSNP在法医实战应用中评估其在DNA与RNA水平的分型一致性提供数据基础和方法参考。
Objective To evaluate the consistency of DNA coding region single nucleotide polymorphism(cSNP)genotyping at the DNA and RNA levels in common body fluid samples based on the next-generation sequencing platform.Methods After extensive literature retrieval,25 cSNP loci of 8 human tissue-specific mRNAs in peripheral blood,semen and vaginal secretion were selected.Two cSNP multiplex genotyping panels based on DNA and RNA,respectively,were developed for use on the MiSeq FGx sequencing platform.45 body fluid samples(including 14 peripheral blood samples,15 semen samples and 16 vaginal secretion samples)were sequenced and analyzed.The inconsistent typing results of DNA and RNA were rechecked by Sanger sequencing.Results The results of cSNP genotyping at the DNA and RNA levels in peripheral blood were completely consistent.Among the 15 semen samples,the genotypes of rs1995640 and rs1995641 on the TGM4 gene were inconsistent in 3 cases.Among the 16 vaginal secretion samples,there were 2 cases,1 case and 2 case with inconsistent results of rs3869098,rs10947121 and rs12110470 in MUC22 gene,respectively.Conclusion In this study,MiSeq FGx sequencing and Sanger sequencing were used to test 25 cSNP loci with body fluid tissue specificity.The same typing results at the DNA and RNA levels were observed at 20 cSNPs.Inconsistent genotypes at the DNA and RNA levels were observed at 5 cSNPs on the TGM4 and MUC22 genes.This study provides experimental methods and data for forensic cSNP studies.
作者
娄丹妮
赵一霞
缪磊
赵杰
张驰
康克莱
胡胜
叶健
王乐
Lou Danni;Zhao Yixia;Miao Lei;Zhao Jie;Zhang Chi;Kang Kelai;Hu Sheng;Ye Jian;Wang Le(School of Forensic Medicine,Shanxi Medical University,Shanxi,Jinzhong 030600,China;Key Laboratory of Forensic Genetics of Ministry of Public Security,Institute of Forensic Science,Ministry of Public Security,Beijing 100038,China)
出处
《中国法医学杂志》
2025年第3期295-301,307,共8页
Chinese Journal of Forensic Medicine
基金
国家重点研发计划课题(2022YFC3341001)
公安部科技强警基础工作计划(2023JC15)
中央级公益性科研院所基本科研业务费(2024JB027)。
关键词
法医遗传学
体液鉴定
二代测序
编码区单核苷酸多态性
Forensic genetics
Body fluid identification
Next-generation sequencing
Coding region single nucleotide polymorphism
