PowerPlex21直扩法与磁珠法用于毛发STR检验的比较

目的比较Power Plex21直接扩增法与磁珠提取后扩增法用于带毛囊毛发STR检验的效果。方法带毛囊毛发平均分成2组,每组含拔取毛发20根、自然脱落带毛囊毛发20根,剪取毛囊后1组用Power Plex21试剂盒直接扩增,1组先用磁珠法提取DNA然后扩增Power Plex21试剂盒,扩增产物在3130XL遗传分析仪上进行电泳检测分析。结果拔取毛发直接扩增和磁珠提取后扩增均成功检出全部STR基因型。自然脱落带毛囊毛发直接扩增检出STR基因型的成功率为55%,磁珠提取后扩增检出STR基因型的成功率为25%。结论 Power Plex21直接扩增法比磁珠提取后扩增更适于带毛囊毛发的STR检验。     

两无关个体单亲亲子鉴定不排除1例

1案例资料2004年4月5日,黄埔分局送来在某医院提取的两名涉嫌被拐卖男童(均约6个月大)的口腔拭子,要求进行DNA检验,以确定是否有血缘关系。用Chelex-100法提取该两名儿童的口腔拭子DNA。先后用Identifiler试剂盒、PowerPlex 16试剂盒、FFFL试剂盒和PowerPlex Y试剂盒进行扩增。扩增产物用AB I 3100型遗传分析仪检测分型,并进行m tDNA L15997~L16401片段序列测定。经Identifiler系统15个常染色体STR基因座检验,两男孩的基因型不同,但每个基因座均至少有1个相同的等位基因,见表1。再使用Power Plex16和FFFL试剂盒检验,在增加的…     

广东潮安地区汉族人群15个STR基因座遗传多态性

本研究对中国广东潮安地区汉族500名健康无关个体进行15个常染色体STR基因座遗传多态性调查。采用Chelex-100法提取样本DNA,用Power Plex16 System试剂盒进行扩增及检测。结果在15个基因座共检测出165个等位基因和620种基因型,其分布符合Hardy-Weinberg平衡定律(P0.05),对所调查的汉族人群,本文15个STR基因座具有较好识别能力。     

云南苗族群体15个STR基因座遗传多态性

本文对中国云南地区苗族219个健康无关个体进行15个常染色体STR基因座遗传多态性调查。采用Chelex-100法提取样本DNA,用Power Plex16 System试剂盒进行扩增及检测。结果在15个STR基因座共检出161个等位基因和438种基因型,其分布符合Hardy-Weinberg平衡定律(P0.05)。所调查的云南人群15个STR基因座具有较好识别能力。     

河南汉族人群23个Y-STR基因座遗传多态性

<正>人类Y染色体是男性特有,呈父系、单倍型遗传特征,在法医学检验中作用独特。本研究采用Power PlexY23荧光标记复合扩增系统(美国Promega公司),对河南汉族人群23个Y-STR基因座的遗传多态性进行调查,旨在为法医学应用及群体遗传学研究提供基础数据。1材料与方法1.1样本1 100例居住于河南、无父系亲缘关系的汉族男性个体的纸质血样取自实验室日常检案积累。     

中国东莞地区汉族人群22个Y-STR基因座的遗传多态性

目的调查22个Y-STR基因座在东莞地区汉族人群中的遗传多态性。方法应用Power Plex?Y-23试剂盒扩增22个基因座,扩增产物经3130XL型遗传分析仪电泳检测,用Gene Mapper软件进行等位基因分型。结果1042名男性共检出1042种不同的基因型,22个Y-STR基因座的GD值为0.413 9~0.959 9。结论 22个Y-STR基因座在东莞地区汉族人群中有较好分布,对法医学应用和人类群体遗传学研究具有重要价值。     

PentaD、D21S11基因座分型异常分析3例

<正>1案例资料1.1简要案情30例身源不明的儿童血样,来源于拐卖儿童的送检检材。在STR分型检验中,发现其中3例(1例男性,2例女性)Penta D及D21S11基因座分型异常。1.2检验方法DNA分型检验分别用Identifiler Plus(Life Technogies公司,美国)、Power Plex21(Promega公司,     

辽南地区汉族群体23个STR基因座遗传多态性

本文对中国辽南地区汉族群体544个无关个体23个STR基因座遗传多态性进行调查。采用磁珠法提取基因组DNA,采用Power PlexFusion System试剂盒进行扩增及检测。结果在22个常染色体STR基因座共检出257种等位基因,其分布符合Hardy-Weinberg平衡定律(P0.05)。累计个体识别率达1-2.3×1026,累计非父排除率达0.999 999 999 98。本文调查的23个STR基因座除4个多态性稍差外,其余均有较好的多态性,具有较高应用价值。     

亲子鉴定中D8S1179基因座等位基因丢失分析1例

1案例资料1.1简要案情田某,女,36岁,因落户需要,要求对其与女儿进行亲权鉴定。1.2检验方法采用Chelex法抽取血样样本DNA;分别用Power Plex21试剂盒（普洛麦格（北京）生物科技有限公司）、GoldeneyeTM20A试剂盒（北京基点认知公司）进行复合PCR扩增;用3130XL遗传分析仪（美国AB公司）进行毛细管电泳,用GeneMapperIDv3．2软件进行基因型分析。     

贵州省汉族群体22个STR基因座遗传多态性

应用Power PlexFusion 5色荧光标记复合扩增系统,对605名贵州省汉族无关个体22个常染色体STR基因座遗传多态性进行调查。结果 22个STR基因座共观察到260个等位基因,945种基因型,各基因座基因型分布均符合Hardy-Weinberg平衡定律(P0.05)。22个基因座遗传学参数值范围分别为:H值0.625~0.929,DP值0.804~0.985,PE值0.322~0.855,PIC值0.57~0.91,累计非父排除概率为0.999 999 999,累计个人识别率为1-1.301 4×10-26。22个STR基因座在贵州汉族人群中具有良好的遗传多态性,具有较高的法医学应用价值。     

3种常用PCR扩增试剂盒检验血样DNA的结果比较

目的探讨常用的ProfilerPlusTM、IdentifilerTM与Powerplex(16试剂盒检验血样DNA的差异。方法510名无关中国汉族个体血样,分别用ProfilerPlusTM与Powerplex(16试剂盒进行DNA检验,然后对有不同检验结果的同一样本,再用ProfilerPlusTM、IdentifilerTM与Powerplex(16等三种试剂盒进行检验,并比较其结果。结果在510名个体血样的DNA检测结果中,发现同一样本有不同结果的有7例,其差异率为1.3725%;ProfilerPlusTM、IdentifilerTM与Powerplex(16各有1例在D13S317或FGA基因座上出现等位基因缺失现象,缺失率为0.1961%;ProfilerPlusTM有5例在D8S1179基因座上出现扩增严重不平衡现象,相应的IdentifilerTM与Powerplex(16试剂盒的检验结果为正常杂合子。结论ProfilerPlusTM、IdentifilerTM与Powerplex(16试剂盒检验血样DNA均会出现扩增不平衡和/或基因丢失现象,其发生几率IdentifilerTM与Powerplex(16试剂盒较ProfilerPlusTM少。     

D8S1179基因座等位基因丢失现象的研究

目的 探讨在进行STR分型时发生的等位基因“丢失”情况的原因。方法 分别采用Profiler Plus试剂盒和GenBank数据库设计的引物,对D8S1179基因座进行基因分型,并对丢失的等位基因进行测序分析。结果D8S1179基因座丢失的等位基因在第147位碱基出现G-A转换。在中国人群中,该位置出现碱基转换的频率是0.50×10-2(在2013个无关个体中观察到10例无关变异事件)。结论 第147位碱基转换正好位于Prfiler Plus试剂盒中D8S1179基因座的引物结合区域,导致扩增时等位基因丢失。     

STR primer concordance study.

Over 1500 population database samples comprising African Americans, Caucasians, Hispanics, Native Americans, Chamorros and Filipinos were typed using the PowerPlex 16 and the Profiler Plus/COfiler kits. Except for the D8S1179 locus in Chamorros and Filipinos from Guam, there were eight examples in which a typing difference due to allele dropout was observed. At the D8S1179 locus in the population samples from Guam, there were 13 examples of allele dropout observed when using the Profiler Plus kit. The data support that the primers used in the PowerPlex 16, Profiler Plus, and COfiler kits are reliable for typing reference samples that are for use in CODIS. In addition, allele frequency databases have been established for the STR loci Penta D and Penta E. Both loci are highly polymorphic.     

Identification of a D8S1179 primer binding site mutation and the validation of a primer designed to recover null alleles

A population study of Chamorros and Filipinos using short tandem repeat (STR) loci amplified with the AmpFlSTR Profiler Plus PCR amplification kit demonstrated an excess of observed homozygosity at the D8S1179 locus. Use of a different set of D8S1179 primers to type the same samples did not demonstrate an excess of homozygosity and showed discordant genotypes at the D8S1179 locus. A single point mutation, G-to-A transition, 16 nucleotides from the 3' end of the reverse primer, was identified to cause allele dropout when using the AmpFlSTR Profiler Plus primer set. An additional D8S1179 reverse primer specific for the variant was constructed resulting in the recovery of the null allele. The primer was included in the newly developed AmpFlSTR Identifiler PCR amplification kit. No deleterious effects or non-specific peaks were observed in validation experiments evaluating primer concentration, Mg2+ concentration, annealing temperature and population samples.     

Profiler Plus试剂盒扩增Amelogenin基因座变异1例

目的探讨日常检案及DNA数据库建设中常用的ProfilerPlus系统扩增法医生物检材可能出现的变异情况。方法分别采用ProfilerPlus试剂盒扩增、毛细管电泳分析及Amelogenin单基因座扩增、银染技术分型两种方法分别检测两份血痕的基因型。结果应用ProfilerPlus试剂盒扩增、毛细管电泳分析发现X-Y同源Amelogenin基因座的X片段丢失,而运用Amelogenin单基因座扩增、银染技术分型则X片段正常出现。结论应用ProfilerPlus系统分析法医生物检材,有可能出现等位基因丢失现象,此现象需要引起我们在日常检案及DNA数据库建设中的足够重视。     

Population data of nine STRs of Mexican-Mestizos from Mexico City

One hundred and thirteen individuals were PCR-typed for nine STR loci with the AmpFlSTR Profiler Plus PCR amplification kit, including the following autosomal STRs: D3S1358, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317 and D7S820. Allele frequencies for each STR were estimated, and they were compared to other populations. Genotype distribution by locus and by two-loci combination was in agreement with Hardy-Weinberg expectations for all nine STRs. For this region of Mexico, the combined probability of exclusion (PE) and power of discrimination (PD) were estimated: PE=99.964% and PD>99.999%.     

Validation of short tandem repeats (STRs) for forensic usage: performance testing of fluorescent multiplex STR systems and analysis of authentic and simulated forensic samples

The amplification and typing conditions for the 13 core CODIS loci and their forensic applicability were evaluated. These loci are CSF1PO, FGA, TH01, TPOX, vWA, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, and D21S11. Results were obtained using the multiplex STR systems AmpFlSTR Profiler Plus and AmpFlSTR COfiler (Applied Biosystems, Foster City, CA), GenePrint PowerPlex (Promega Corporation, Madison, WI), and subsets of these kits. For detection of fluorescently labeled amplified products, the ABI Prism 310 Genetic Analyzer, the ABI Prism 377 DNA Sequencer, the FMBIO II Fluorescent Imaging Device, and the Fluorlmager were utilized. The following studies were conducted: (a) evaluation of PCR parameter ranges required for adequate performance in multiplex amplification of STR loci, (b) determination of the sensitivity of detection of the systems, (c) characterization of non-allelic PCR products, (d) evaluation of heterozygous peak intensities, (e) determination of the relative level of stutter per locus, (f) determination of stochastic PCR thresholds, (g) analysis of previously typed case samples, environmentally insulted samples, and body fluid samples deposited on various substrates, and (h) detection of components of mixed DNA samples. The data demonstrate that the commercially available multiplex kits can be used to amplify and type STR loci successfully from DNA derived from human biological specimens. There was no evidence of false positive or false negative results and no substantial evidence of preferential amplification within a locus. Although at times general balance among loci labeled with the same fluorophore was not observed, the results obtained were still valid and robust. Suggested criteria are provided for determining whether a sample is derived from a single source or from more than one contributor. These criteria entail the following: (a) the number of peaks at a locus, (b) the relative height of stutter products, and (c) peak height ratios. Stochastic threshold levels and the efficiency of non-templated nucleotide addition should be considered when evaluating the presence of mixtures or low quantity DNA samples. Guidelines, not standards, for interpretation should be developed to interpret STR profiles in cases, because there will be instances in which the standards may not apply. These instances include (a) a primer binding site variant for one allele at a given locus, (b) unusually high stutter product, (c) gene duplication, and (d) translocation.     

Genetic data of 9 STR loci from Henan Province (central China)

Allele frequencies of the nine short tandem repeats (STR) loci D8S1179, D21S11, D18S51, D5S818, D3S1358, D7S820, vWA, FGA (AmpFlSTR Profiler Plus) were determined in a population sample of unrelated individuals living in central China.     

Portuguese population and paternity investigation studies with a multiplex PCR--the AmpFlSTR Profiler Plus

A Portuguese Caucasian population of 146 unrelated individuals was studied. DNA samples were amplified by multiplex PCR for D3S1358, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317 and D7S820 using the AmpFlSTR Profiler Plus PCR Amplification Kit (Perkin-Elmer). All loci met Hardy-Weinberg expectations. Forensic statistical parameters were according to those obtained by other authors. Statistical differences were observed concerning three loci when comparing the Portuguese Caucasian population and an Italian Caucasian population, although these differences mainly concern the less frequent alleles. Eighty-three paternity investigation cases were analysed. Exclusions in between three and nine loci were observed in all the 23 exclusion cases obtained. Most of the non-exclusion cases had probability of paternity > 99.9%. Two cases with an isolated genetic incompatibility between the alleged father and the child were detected, which may indicate probable mutation cases. These results demonstrate that the AmpFlSTR Profiler Plus is a suitable multiplex for paternity investigation in the Portuguese population.     

Allele frequencies for nine STR loci in a Korean ethnic group from northeast China

Allele frequencies for the nine tetrameric short tandem repeats (STR) loci D3S1358, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820 (AmpFlSTR) Profiler Plus PCR Amplification kit, PE Applied Biosystems) were determined in 109 unrelated Korean ethnic group individuals from northeast China.