法医学杂志

法医学杂志 ›› 2009, Vol. 0 ›› Issue (5): 337-340+.

• 论文 • 上一篇    下一篇

甲醛固定石蜡包埋组织STR分型检测

柳燕;李莉;赵珍敏;张素华;赵书民;   

  1. 司法部司法鉴定科学技术研究所上海市法医学重点实验室;苏州大学医学部法医学系;
  • 发布日期:2009-10-25 出版日期:2009-10-28

STR Genotyping in Unbuffered Formalin Fixed Paraffin Embedded Tissue

LIU YAN1,LI LI1,ZHAO ZHEN-MIN1,ZHANG SU-HUA1,2,ZHAO SHU-MIN1(1.SHANGHAI KEY LABORATORY OF FORENSIC MEDICINE,INSTITUTE OF FORENSIC SCIENCE,MINISTRY OF JUSTICE,P.R.CHINA,SHANGHAI 200063,CHINA;2.DEPARTMENT OF FORENSIC MEDICINE,MEDICAL COLLEGE OF SOOCHOW UNIV   

  • Online:2009-10-25 Published:2009-10-28

被引次数

17

摘要： 目的评估10%甲醛固定石蜡包埋组织STR分型结果的影响因素。方法采用QIAGEN法、IQ法、Chelex法对2具新鲜尸体在尸检时常规制备的心、脑、肝、脾、肾、肺、胃、肠石蜡包埋组织进行DNA提取,用AmpFlSTR Identifiler试剂盒进行PCR扩增,在3100-Avant上完成片段分析。另外对15个案例中室温保存1～5年的心、肝、肺、肠存档石蜡包埋组织共56份采用同样的方法进行STR分型。以STR基因座检出率评估分型有效性。结果各种组织DNA片段均随着保存时间延长而持续降解,其中心、肺组织STR基因座检出率与保存时间存在线性相关。相同保存时间时,各种组织基因座检出率差异有统计学意义。其中以肺组织在不同保存时间中基因座检出率最高。结论在甲醛固定时间一定的条件下,存放时间、组织类型、DNA提取方法和PCR模板质量浓度是影响石蜡包埋组织STR分型的重要因素。

关键词: 法医遗传学, 石蜡包埋, STR分型, 影响因素

Abstract: Objective To assess the influential factors of STR genotyping in 10% unbuffered formalin fixed paraffin embedded samples.Methods Eight kinds of autopsy samples including heart,brain,liver,spleen,kidney,lung,stomach and intestine tissue from 2 corpse were fixed with 10% unbuffered formalin and embedded with paraffin according to the routine procedure from which the DNA were extracted with three different methods(QIAGEN,IQ and Chelex).STR profile were analyzed with AmpFlSTR Identifiler Kit and capillary electrophoresis on genetic analyzer 3100-Avant.STR profiles of 56 archival paraffin embedded samples from 15 cases were also analyzed with methods as mentioned.These archival samples,including heart,liver,lung and intestine tissue,had been preserved for 1 to 5 years in ambient temperature.Effectiveness of STR genotyping was assessed with the recalling ration of the 15 STR loci composing of the Identifiler Kit.Results Significant difference of the recalling ration was statistically revealed among the different types of paraffin embedded sample with same preserving period.Moreover,the STR recalling ration was continuously lowering with the prolongation of preserving period in all of the samples.The linear relation-ship between the STR recalling ratio and the preserving period was showed in lung and heart sample.The STR recalling ration in lung sample was higher than that in the other types of paraffin embedded sample.Conclusion Preserving period,tissue type,extracting method of DNA and the PCR template concentration were the most important influential factors for successfully STR genotyping paraffin embedded samples,which fixed with unbuffered formalin for the same time.

Key words: forensic genetics, paraffin embedding, STR typing, influential factors