期刊文献+

荧光标记复合扩增检测4个STR基因座多态性 被引量:1

Study on the polymorphisms of four STR loci using fluorescence-labeled multiplex-PCR technique
在线阅读 下载PDF
导出
摘要 目的建立荧光标记复合扩增D1S2142,D13S1492,D14S306,D15S659基因座检测分型方法,并对成都汉族群体4个基因座的遗传多态性进行调查。方法用6-FAM标记D1S2142和D15S659引物,HEX、TMR分别标记D14S306和D13S1492引物,PCR复合扩增,310基因分析仪电泳自动收集电泳结果数据,GeneScan Analysis Software3.7NT软件计算扩增产物片段相对大小,Genotyper(3.7NT软件进行样本基因型分型,建立了荧光标记复合扩增检测4个STR基因座基因型的方法,对145名成都汉族无关个体样本进行分型。结果荧光标记复合扩增D1S2142,D13S1492,D14S306,D15S659基因座,每个STR基因座都获得了清晰的基因型分型结果。145份样本,4个STR基因座分别检出10,14,7,12个等位基因和22,54,21,39种基因型,其基因型分布均符合Hardy-W e inberg平衡。4个基因座在成都汉族群体的杂合度分别依次为0.7793,0.8345,0.7793和0.8345;多态信息含量分别依次为:0.7656,0.8730,0.7470和0.8312。累计非父排除率为0.9783,累计个人识别机率为0.9999 917。结论荧光标记复合扩增D1S2142,D13S1492,D14S306,D15S659基因座,可实现对每个基因座准确分型;成都汉族群体该4个基因座的遗传学数据,可为群体遗传学和法医学研究与应用提供基础资料。 Objective To establish a method for typing STR loci D1S2142, D13S1492, D14S306 and D15S659 by fluorescence - labeled multiplex - PCR technique, by which the genetic data of Han population in Chengdu area were obtained. Methods Primers of STR loci D1S2142 and D15S659 were labeled with 6 - FAM, and primer of D14S306 and D13S1492 were labeled with HEX and TMR respectively. Multiplex - PCR technique was applied to amplifying the target DNA. The electrophoresis of PCR products was accomplished on the 310 Genetic Analyzer with the sample's electrophoresis data collected automatically using Data Collection Software 3.0. The relativeneScan Analysis Software 3.7NT and genotypedfragment lengths of PCR products were calculated by the Geby the Genotyper 3.7NT Software. Therefore, a method of multiplex - PCR labeled with fluorescence to detect the genotypes of the four STR loci had been established. We used this method to investigate the genotype distributions and allele frequencies in 145 EDTA - blood specimens collected from unrelated individuals in Chengdu Han population. Results Four STR loci were exactly determined by fluorescence - labeled multiplex - PCR technique. Each locus was successfully genotyped in all 145 samples. 10, 14, 7, 12 alleles and 22, 54, 21, 39 genotypes were observed in D1S2142, D13S1492, D14S306, and D15S659 locus, respectively. The distributions of genotype for four STR loci in Chengdu Han population were in accordance with Hardy - Weinberg equilibrium. The heterozygotes and polymorphism information component ( PIC ) of the four STR loci were 0.7793, 0.8345, 0. 7793, 0. 8345 and 0. 7656, 0. 8730, 0. 7470, 0. 8712 respectively. The combined exclusion probability and the combined discrimination power of the four STR loci in Chengdu Han population were 0.9783 and 0.9999917. Conclusion The STR loci D1S2142, D13S1492, D14S306, and D15S659 could be correctly genotyped using fluorescence - labeled multiplex - PCR technique. The population genetic information of the four STR loci may be used as the basic data in the study and application in genetics and forensic practice.
出处 《中国法医学杂志》 CSCD 2005年第4期218-221,共4页 Chinese Journal of Forensic Medicine
基金 四川省科技厅应用基础项目基金(03JY029-074-1) 纽约中华医学基金(CMB 2000-636)。
关键词 法医物证学 短串联电复 复合扩增 基因座 基因频率 Forensic biological evidence Short tandem repeats(STR) Multiplex polymerase chain reaction Locus Allelic frequencies
  • 相关文献

参考文献6

  • 1Krenke BE, Tereba A, Anderson S J, et al. Validation of a 16 -locus fluorescent multiplex system [J]. J Forensic Sci,2002, 47(4) :773 -785.
  • 2Micka KA, Amiott EA, Hockenberry TL, et al. TWGDAM validation of a nine - locus and a four - locus fluorescent STR multiplex system [J]. J Forensic Sci, 1999, 44(6):1243 - 1257.
  • 3Walsh PS, Metzger DA, Higuchi R. Chelex 100 as a medium for simple extraction of DNA for PCR - based typing from forensic material[ J]. Biotechniwues, 1991, 10: 506 - 513.
  • 4Bar W, Brinkmann B, Budowle B, et al. DNA recommendations. Further report of the DNA Commission of the ISFH regarding the use of short tandem repeat systems. International Society for Forensic Haemogenetics [ J ]. Int J Legal Med, 1997, 110(4):175-176.
  • 5Hou Y, Prinz M, Staak M. Comparison of different tests for deviation from Hardy - Weinberg equilibrium of AMFLP population data[M]. In: Bar W, Fiori, Ross UR eds. Advance in forensic haemogenetics 5. Berlin: Springer - Verlag,1994. 511-514.
  • 6邓志辉,吴国光,程良红,魏天莉,苏宇清,郝桂琴.13个STR基因座在亲子鉴定案例中的基因突变观察[J].中国法医学杂志,2003,18(3):150-153. 被引量:21

二级参考文献12

  • 1Toshimichi Yamamoto, Keiji Tamaki, Xiu-Lin Huang, et al. The application of minisatellite variant repeat mapping by (MVR-PCR) in a paternity case showing false exclusion due to STR mutation [J]. J Forensic Sci, 2001; 46 (2): 374
  • 2刘秋玲 伍新尧 吕德坚 等.亲子鉴定中2个STR基因突变1例[J].中国法医学杂志,2001,16(4):238-238.
  • 3Annual Report Summary for 2000, Prepared by the Parentage Testing Standards Committee. (http: //www. aabb. Org/ about the aabb/stds and accred/ptannrpt00, pdf)
  • 4Walsh PS, Metzger DA, Higuchi R. Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material [J]. Biotechniques, 1991, 10:506
  • 5Lee HS, Lee JW, Han GR, et al. Motherless case in paternity testing [J]. Forensic Sci Int, 2000, 114:57
  • 6Gjertson DW. The effect of an isolated single-locus inconsistency in the statistical evaluation of paternity [M]. In: Walker RH. Paternity testing accreditation requirements manual of the American Association of Blood Banks. 3rd ed. San Francisco: AABB press, 1998. 9
  • 7Weber JL, Wong C. Mutation of human short tandem repeats. Hum Molec [J]. Genet, 1993, 2:113
  • 8Levinson G, Gutman GA. Slipped strand mispriming: a major mechanism for DNA sequence evolution [J]. Mol Biol Evol, 1987, 4:203
  • 9Annual Report Summary for Testing in 2001, Prepared by the Parentage Testing Program Unit [Z]. http: // www. aabb. Org/about the aabb/stds and accred/ptannrpt01. pdf
  • 10Bernd B, Michael K, Franz N, et al. Mutation rate in human microsatellites: influence of the structure and length of the tandem repeat [J]. Am J Hum Genet, 1998, 62: 1408

共引文献20

同被引文献4

  • 1黄代新,朱传红,方慧,刘翠兰,杨罕,杨庆恩.武汉汉族群体3个多拷贝Y-STR基因座的遗传多态性[J].中国法医学杂志,2007,22(2):76-80. 被引量:7
  • 2Gill P, Brenner C, Brinkmann B, et al. DNA commission of the International Society of Forensic Genetics: recommendations on forensic analysis using Y-chromosome STRs [ J ]. Int J Leg Med, 2001, 114:305-309.
  • 3Park M J, Lee H Y, Yoo J E, et al. Forensic evaluation and haplotypes of 19 Y-chromosomal STR loci in Koreans [ J]. Forensic Sci Int, 2005, 152 : 133-147.
  • 4Hedman M V, Pimenoff M, Lukka P, et al. Analysis of 16 Y-STR loci in the Finnish population reveals a local reduction in the diversity of male lineages [ J ]. Forensic Sci Int, 2004, 142:37-43.

引证文献1

二级引证文献2

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部