Analysis of AmpFlSTR MiniFiler™ loci and its forensic application

The allele frequencies of eight MiniFiler™ loci have been analyzed in 101 Japanese individuals living in Kanagawa with informed consent by means of ABI 310 Genetic Analyzer. A total of 7 alleles for D13S317, 8 alleles for D7S820, 11 alleles for D2S1338, 11 alleles for D21S11, 5 alleles for D16S539, 14 alleles for D18S51, 8 alleles for CSF1PO, and 13 alleles for FGA were observed. The polymorphic profiles of these MiniFiler™ loci in the present study were essentially the same as those obtained by using the AmpFlSTR^® Identifiler^® PCR Amplification kit. The combined matching probability of eight MiniFiler™ loci and cumulative probability of paternity exclusion were estimated as 1.97 × 10⁻¹⁰ and 0.9996, respectively. The MiniFiler™ kit was useful for individual identification in forensic analysis.     

Analysis of DNA profiles extracted from degraded samples from archival of formalin fixed tissue included in paraffin (FFTIP) and hairs

The possibility of studying DNA extracted from archival of formalin fixed tissue included in paraffin (FFTIP) enables valuable retrospective investigations. However, according to some authors it is difficult to obtain genomic DNA of good quality, since the process of fixation often results in fragmentation of DNA. In order to evaluate the quality and quantity of DNA extracted, necropsy samples of FFTIP (spleen/lung) and hairs, with or without bulbs, were analyzed using three methods of extraction (QIAamp DNA mini, QIAamp DNA micro-kit and phenol–chloroform followed by microcon YM-30). The amount of DNA recovered was quantified by spectrophotometer. The β-actin, amelogenin gene and the profiles of STR were analyzed. Based on experimental results, a general guideline concerning the appropriate extraction method according to the tissue and the quantity of the starting material for the analysis of DNA from FFTIP and hairs could be suggested.     

Heteroplasmies detected in an amplified mitochondrial DNA control region from a small amount of template

When mitochondrial DNA (mtDNA) heteroplasmies are detected, they often confound forensic identification, especially if they are the result of poor biological sampling. In this study, we determined the ratio of heteroplasmy in samples that were amplified from a very small amount of template mtDNA or a few cells using a highly sensitive nested polymerase chain reaction (PCR) procedure and a direct sequencing analysis. As a result, more than half of the detected sequences (i.e., 17/20, 15/20, and 14/20) showed homoplasmy derived from a variation in the heteroplasmy proportion when only 10 copies of template mtDNA samples were amplified and analyzed. Additionally, with products amplified from one or several white blood cells (WBCs), several previously undetected heteroplasmies were detected. These results indicate the risks associated with using highly sensitive mtDNA techniques in forensic investigations because of the variable proportions of heteroplasmy or nucleotide substitutions that can possibly be detected from a very small biological sample.     

一种用于降解DNA样本的性别鉴定方法

目的 建立一种采用PCR技术对降解DNA样本进行性别鉴定的新方法。方法 采用针对amelogenin基因X染色体外显子3bp缺失设计的引物AMELU1及AMELD1,对在室温环境下放置5－15年的男、女血痕标本各50例、毛发各20例、骨骼各20例以及现场提取5－－20天的男、女腐败肌肉各10例标本中提取的降解DNA样本进行扩增。用PGA（9％T,3％C）电泳、银染显带检测扩增产物。结果 所有样本均得到正确结果,男性检材表现为83bp的Y特异性及80bp的X特异性2条谱带,而女性检材仅有1条80bp的X特异性谱带。结论 用针对amelogenin基因X染色体外显子3bp缺失设计的引物AMELU1及AMELD1鉴定性别的方法灵敏、可靠、方便,是降解DNA检材性别鉴定十分理想的方法。     

一种用于降解DNA样本的性别鉴定方法

目的　建立一种采用PCR技术对降解DNA样本进行性别鉴定的新方法。　方法　采用针对amelogenin基因X染色体外显子 3bp缺失设计的引物AMELU1及AMELD1,对在室温环境下放置 5～ 15年的男、女血痕标本各 5 0例、毛发各 2 0例、骨骼各 2 0例以及现场提取 5 - - 2 0天的男、女腐败肌肉各 10例标本中提取的降解DNA样本进行扩增。用PAG( 9%T ,3 %C)电泳、银染显带检测扩增产物。　结果　所有样本均得到正确结果 ,男性检材表现为 83bp的Y特异性及 80bp的X特异性 2条谱带 ,而女性检材仅有 1条 80bp的X特异性谱带。　结论　用针对amelogenin基因X染色体外显子 3bp缺失设计的引物AMELU1及AMELD1鉴定性别的方法灵敏、可靠、方便 ,是降解DNA检材性别鉴定十分理想的方法。     

CSF1PO,TPOX和TH01基因座复合扩增及其在苗族人群中的基因频率分布

在同一反应体系中，对CSFIPO，TPOX和TH01三个STR基因座做复合扩增，采用高分辨率的聚丙烯酸胺凝胶电泳分离、银染法显影技术，对云南苗族的3个基因座基因频率分布进行调查。CSFIPO基因座，观察到8个等位基因，17个基因型；TPOX基因座，观察到6个等位基因，14个基因型；TH01基因座，观察到6个等位基因，19个基因型。     

ABO基因分型及其在法医学中的应用

为建立一种ABO血型系统基因分型方法，采用PCR-RFLP技术，成功地将ABO系统区分为AA，AO，AB，OO，BB，BO六种基因型。对240名中国汉族无关个体血样的ABO（基因型频率调查结果表明，6种基因型的频率分布为0．0125～0．3834，符合Hardy－Weinbeng遗传平衡法则（P＞0．1），其DP值为0．8161。家系分析表明，亲代a、b、o基因传递遵守孟德尔遗传规律。对法医学中常见的血痕、混合斑、骨组织及毛发根部等生物样品进行检测，均能准确判定ABO基因型，并可在实际案件鉴定中应用。     

浙江汉族人群6个STR基因座的遗传多态性的调查

目的进一步完善浙江省汉族人群STR基因座遗传多态性的调查,为其应用提供基础数据。方法采用AmpF1STRSGMplus和AmpF1STRCoficer反应试剂盒,使用ABI310型基因分析仪对浙江汉族人群200名无关个体血样进行了D16S539、D2S1338、D19S433、TH01、TPOX和CSF1PO6个STR基因座遗传学分析。结果分别发现了9、15、15、11、8、10个等位基因,发现的基因型分别为23、42、35、19、16、17个,其分布经X2检验均符合Hardy-Weinberg平衡定律,并分别统计了6个STR基因座的H、DP、PM、PE及PIC参数。结论6个STR基因座适合法医学应用。     

PCR法对HLA-DQ_α基因的分型及其在性犯罪鉴定中的应用

本文应用 PCR 法及 ASO 探针斑点杂交技术,对100例无关个体血液 DNA 及10例性犯罪案件混合斑中精子 DNA 进行了 HLA-DQα基因的扩增及其 DNA 分型。结果正常人0.1～0.3μgDNA 就能满足 DQα基因扩增的需要。在100例个体中可以观察到由4种等位基因组成的10种 DQα基因型。10例不同条件的混合斑中精子 DNA 经30～60次扩增后与 ASO 探针杂交均能准确地判定 DQα基因型。HLA-DQα是个体识别能力较强的遗传标志。本文为性犯罪中精子来源的个体识别提供了一个新方法,具有一定的实用价值。     

广西地区3个群体15个STR基因座频率多态性

目的调查广西地区仡佬族、仫佬族、瑶族无关个体的15个STR基因座(D8S1179、1321S11、D7S820、CSF1PO、D3S1358、TH01、D13S317、D16S539、D2S1338、D19S433、vWA、TPOX、D18S51、D5S818、FGA)多态性,研究其在法医学检验中的应用价值。方法 应用AmpFISTR IdentifilerTM荧光标记复合扩增系统,对314例仡佬族、332例仫佬族、238例瑶族无关个体血样DNA进行15个STR基因座的复合扩增,用ABI 3100遗传分析仪对扩增产物进行检测,GeneScan、GenoTyper软件进行基因分型,统计计算15个STR基因座的群体遗传学参数。结果IdertifilerTM系统的15个STR基因座在仡佬、仫佬、瑶族的累积偶合率为1.839×10-16~5.073×10-17,累积非父排除率为0.9999983~0.9999991。结论该15个STR基因座足可满足仡佬族、仫佬族、瑶族法医学的个体识别及亲权鉴定的需要。