This work is aimed at describing the proceedings and parameters used to validate PowerPlex■ Fusion 6C System,the polymerase chain reaction (PCR) amplification kit by Promega,for posterior implementation in the labora...This work is aimed at describing the proceedings and parameters used to validate PowerPlex■ Fusion 6C System,the polymerase chain reaction (PCR) amplification kit by Promega,for posterior implementation in the laboratorial routine of the Forensic Genetic Service.The PowerPlex■ Fusion 6C System allows multiplex PCR,through simultaneous amplification and posterior detection by fluorescence of 27 loci.Characterization of the kit was made according to the laboratory's internal validation procedure based on validation guidelines from Scientific Working Group on DNA Analysis Methods.Some parameters were evaluated,such as specificity,analytical thresholds,sensitivity,precision,mixture studies,DNA control samples,a proficiency test and changes in the PCR-based procedures: final reaction volume and cycle number,changes in the reaction mixture for direct amplification.This kit proved to be very robust and the results are in concordance with previous developmental validation by the manufacturer.In some parameters,the results were better than expected.展开更多
Insertion/deletion polymorphisms(InDels)have been treated as a prospective and helpful genetic marker in the fields of forensic human identification,anthropology and population genetics for the past few years.In this ...Insertion/deletion polymorphisms(InDels)have been treated as a prospective and helpful genetic marker in the fields of forensic human identification,anthropology and population genetics for the past few years.In this study,we developed a six-dye multiplex typing system consisting of 34 autosomal InDels and Amelogenin for forensic application.The contained InDels were specifically selected for Chinese population with the MAF≥0.25 in East Asia,which do not overlap with the markers of Investigator^(■)DIPplex kit.The typing system was named as GoldeneyeTM DNA ID System 35InDel Kit,and a series of developmental validation studies including repeatability/reproducibility,concordance,accuracy,sensitivity,stability,species specificity and population genetics were conducted on this kit.We confirmed that the 35InDel kit is precise,sensitive,species specific and robust for forensic practice.Moreover,the 35InDel kit is capable of typing DNA extracted from forensic routine case-type samples as well as degraded samples and mixture samples.All markers are proved to be highly polymorphic with an average observed heterozygosity(He)of 0.4582.The combined power of discrimination(CPD)is 0.999999999999978 and the combined power of exclusion in duos(CPE_(D))and trios(CPE_(T))are 0.978837 and 0.999573,respectively,which are higher than those of the Investigator^(■)DIPplex kit.Thus,the GoldeneyeTM DNA ID System 35InDel kit is suitable for forensic human identification and could serve as a supplementary typing system for paternity testing.展开更多
This review summarizes the scientific basis of forensic gait analysis and evaluates its use in the Netherlands,United Kingdom and Denmark,following recent critique on the admission of gait evidence in Canada.A useful ...This review summarizes the scientific basis of forensic gait analysis and evaluates its use in the Netherlands,United Kingdom and Denmark,following recent critique on the admission of gait evidence in Canada.A useful forensic feature is(1)measurable,(2)consistent within and(3)different between individuals.Reviewing the academic literature,this article found that(1)forensic gait features can be quantified or observed from surveillance video,but research into accuracy,validity and reliability of these methods is needed;(2)gait is variable within individuals under differing and constant circumstances,with speed having major influence;(3)the discriminative strength of gait features needs more research,although clearly variation exists between individuals.Nevertheless,forensic gait analysis has contributed to several criminal trials in Europe in the past 15 years.The admission of gait evidence differs between courts.The methods are mainly observer-based:multiple gait analysts(independently)assess gait features on video footage of a perpetrator and suspect.Using gait feature databases,likelihood ratios of the hypotheses that the observed individuals have the same or another identity can be calculated.Automated gait recognition algorithms calculate a difference measure between video clips,which is compared with a threshold value derived from a video gait recognition database to indicate likelihood.However,only partly automated algorithms have been used in practice.We argue that the scientific basis of forensic gait analysis is limited.However,gait feature databases enable its use in court for supportive evidence with relatively low evidential value.The recommendations made in this review are(1)to expand knowledge on inter-and intra-subject gait variabilities,discriminative strength and interdependency of gait features,method accuracies,gait feature databases and likelihood ratio estimations;(2)to compare automated and observer-based gait recognition methods;to design(3)an international standard method with known validit展开更多
The Investigator 24plex QS Kit(QIAGEN,Hilden,Germany)is a 6-dye fluorescent chemistry short tandem repeat(STR)polymerase chain reaction(PCR)amplification system that simultaneously amplifies 20 of the expanded Combine...The Investigator 24plex QS Kit(QIAGEN,Hilden,Germany)is a 6-dye fluorescent chemistry short tandem repeat(STR)polymerase chain reaction(PCR)amplification system that simultaneously amplifies 20 of the expanded Combined DNA Index System(CODIS)core STR loci,SE33,DYS391,and the standard sex-determining locus,amelogenin,as well as two special internal performance quality sensor controls(QS1 and QS2),which are included in the primer mix to check the PCR performance.This study was designed to be a pilot evaluation of this STR-PCR kit in a Chinese Han population regarding the PCR conditions,sensitivity,precision,accuracy,repeatability,reproducibility,and concordance;tolerance to PCR inhibitors;applicability to real“forensic-type”samples;species specificity;mixture,balance and stutter analyses,and utility in a population investigation.The exhaustive validation studies demonstrated that the Investigator 24plex QS system is accurate,sensitive and robust for STR genotyping.In addition,these genetic markers in the population data in our study indicated that they can also be useful for forensic identification and paternity testing in the Chinese Han population.展开更多
目的对构建的38重InDel快速族群推断体系进行优化,根据DNA分析方法科学工作组(Scien⁃tific Working Group on DNA Analysis Method,SWGDAM)指南进行性能验证,并对其应用于东亚、欧洲、非洲及其混合人群族群推断的准确性进行验证。方法以...目的对构建的38重InDel快速族群推断体系进行优化,根据DNA分析方法科学工作组(Scien⁃tific Working Group on DNA Analysis Method,SWGDAM)指南进行性能验证,并对其应用于东亚、欧洲、非洲及其混合人群族群推断的准确性进行验证。方法以DNA标准品9947A为模板,通过调整引物平衡性、Mg2+终浓度、优化PCR热循环参数和扩增体积等建立最优扩增条件,比较样本的等位基因丢失、非特异性扩增以及推断样本来源是否与已知信息匹配,评价该体系的相关性能。结果本体系的最佳模板用量为0.125~2 ng DNA,InDel分型结果准确,扩增均衡性好,种属特异性好;对混有血红蛋白(≤80μmol/L)、靛蓝(≤40 mmol/L)、钙离子(≤1.0 mmol/L)以及腐殖酸(≤90 ng/μL)等抑制剂的样本具有一定耐受性;可以直接扩增检测血卡、唾液卡,且族群推断结果准确;能够区分两样本的混合DNA样本;对实际案例常见生物检材的检验结果良好。结论38重InDel快速族群推断体系分型结果准确可靠,体系性能符合SWGDAM指南要求,可以准确推断未知个体的非洲、欧洲、东亚及欧亚混合人群来源,可用于法医鉴定实践。展开更多
This article describes a newly devised autosomal short tandem repeat(STR)multiplex polymer-ase chain reaction(PCR)system for 19 autosomal loci(D12S391,D13S317,D16S539,D18S51,D19S433,D2S1338,D21S11,D3S1358,D5S818,D6S10...This article describes a newly devised autosomal short tandem repeat(STR)multiplex polymer-ase chain reaction(PCR)system for 19 autosomal loci(D12S391,D13S317,D16S539,D18S51,D19S433,D2S1338,D21S11,D3S1358,D5S818,D6S1043,D7S820,D8S1179,CSF1PO,FGA,TH01,TPOX,vWA,Penta D and Penta E),27 Y-chromosome STR loci(DYS19,DYS385,DYS3891,DYS38911,DYS390,DYS391,DYS392,DYS393,DYS437,DYS438,DYS439,DYS448,DYS449,DYS456,DYS458,DYS460,DYS481,DYS518,DYS533,DYS570,DYS576,DYS635,DYS627,YGATAH4 and DYF387S1)and amelogenin with six-colour fluorescent labelling.Various parameters were evaluated,such as its accuracy,sensitivity,specificity,stability,ability to ana-lysis of mixtures and effects of changes in the PCR-based procedures.All of the 47 selected STR loci were accurately and robustly amplified from 282 bloodstain samples.The species-spe-cificity was high and some ability to inhibit Hematin was identified.The lowest detectable DNA amount was ≥0.125 ng.All of the male loci of the secondary component were revealed precisely when the control DNA was mixed at male/female and male/male ratios of 1:4 or more.We conclude that the present 19-plex autosomal STR and 27 Y-STR assay is both accur-ate and sensitive.It constitutes an additional powerful tool for forensic applications.展开更多
The EX 16+22Y system is a polymerase chain reaction(PCR)-based amplification kit that enables typing of 15 autosomal short tandem repeat(STR)loci(i.e„D3S1358,D13S317,D7S820,D16S539,TPOX,TH01,D2S133&CSF1PO,D19S433,...The EX 16+22Y system is a polymerase chain reaction(PCR)-based amplification kit that enables typing of 15 autosomal short tandem repeat(STR)loci(i.e„D3S1358,D13S317,D7S820,D16S539,TPOX,TH01,D2S133&CSF1PO,D19S433,vWA,D18S51,D21S11,D8S1179,D5S81&and FGA)and 22 widely used Y chromosome STR(Y-STR)loci(DYS391,DYS527a/b,DYS635,DYS458,DYS456,DYS385a/b,DYS43&DYS44&DYS437,DYS19,DYS576,DYS533,DYS393,DYS389I/n,DYS439,DYS392,Y_GATA_H4,DYS390,and DYS481)which contains 20 core Y-STR recommended by the Ministry of Public Security and amelogenin.This multiplex system was designed for the simultaneous analysis of amelogenin-Y allele mutation,single-source searches,kinship(including familial searching),mixture profiles,international data sharing,and other forensic applications.In this study,the multiplex system was validated for sensitivity of detection,species specificity,DNA mixtures,stability,sizing precision,stutter,reproducibility,and PCR-based conditions according to the Scientific Working Group on DNA Analysis Methods developmental validation guidelines and Chinese criteria for the human fluorescent STR multiplex PCR reagent.The results show that the EX16+22Y system is a robust and reliable amplification kit which can be used for human identification testing.展开更多
文摘This work is aimed at describing the proceedings and parameters used to validate PowerPlex■ Fusion 6C System,the polymerase chain reaction (PCR) amplification kit by Promega,for posterior implementation in the laboratorial routine of the Forensic Genetic Service.The PowerPlex■ Fusion 6C System allows multiplex PCR,through simultaneous amplification and posterior detection by fluorescence of 27 loci.Characterization of the kit was made according to the laboratory's internal validation procedure based on validation guidelines from Scientific Working Group on DNA Analysis Methods.Some parameters were evaluated,such as specificity,analytical thresholds,sensitivity,precision,mixture studies,DNA control samples,a proficiency test and changes in the PCR-based procedures: final reaction volume and cycle number,changes in the reaction mixture for direct amplification.This kit proved to be very robust and the results are in concordance with previous developmental validation by the manufacturer.In some parameters,the results were better than expected.
基金This study was supported by grants from the National Youth Top-Notch Talent of Ten Thousand Program[grant number WRQB2019]the Youth Science and Technology Innovation Leader of Ten Thousand Program[grant number 2018RA2102],China.
文摘Insertion/deletion polymorphisms(InDels)have been treated as a prospective and helpful genetic marker in the fields of forensic human identification,anthropology and population genetics for the past few years.In this study,we developed a six-dye multiplex typing system consisting of 34 autosomal InDels and Amelogenin for forensic application.The contained InDels were specifically selected for Chinese population with the MAF≥0.25 in East Asia,which do not overlap with the markers of Investigator^(■)DIPplex kit.The typing system was named as GoldeneyeTM DNA ID System 35InDel Kit,and a series of developmental validation studies including repeatability/reproducibility,concordance,accuracy,sensitivity,stability,species specificity and population genetics were conducted on this kit.We confirmed that the 35InDel kit is precise,sensitive,species specific and robust for forensic practice.Moreover,the 35InDel kit is capable of typing DNA extracted from forensic routine case-type samples as well as degraded samples and mixture samples.All markers are proved to be highly polymorphic with an average observed heterozygosity(He)of 0.4582.The combined power of discrimination(CPD)is 0.999999999999978 and the combined power of exclusion in duos(CPE_(D))and trios(CPE_(T))are 0.978837 and 0.999573,respectively,which are higher than those of the Investigator^(■)DIPplex kit.Thus,the GoldeneyeTM DNA ID System 35InDel kit is suitable for forensic human identification and could serve as a supplementary typing system for paternity testing.
文摘This review summarizes the scientific basis of forensic gait analysis and evaluates its use in the Netherlands,United Kingdom and Denmark,following recent critique on the admission of gait evidence in Canada.A useful forensic feature is(1)measurable,(2)consistent within and(3)different between individuals.Reviewing the academic literature,this article found that(1)forensic gait features can be quantified or observed from surveillance video,but research into accuracy,validity and reliability of these methods is needed;(2)gait is variable within individuals under differing and constant circumstances,with speed having major influence;(3)the discriminative strength of gait features needs more research,although clearly variation exists between individuals.Nevertheless,forensic gait analysis has contributed to several criminal trials in Europe in the past 15 years.The admission of gait evidence differs between courts.The methods are mainly observer-based:multiple gait analysts(independently)assess gait features on video footage of a perpetrator and suspect.Using gait feature databases,likelihood ratios of the hypotheses that the observed individuals have the same or another identity can be calculated.Automated gait recognition algorithms calculate a difference measure between video clips,which is compared with a threshold value derived from a video gait recognition database to indicate likelihood.However,only partly automated algorithms have been used in practice.We argue that the scientific basis of forensic gait analysis is limited.However,gait feature databases enable its use in court for supportive evidence with relatively low evidential value.The recommendations made in this review are(1)to expand knowledge on inter-and intra-subject gait variabilities,discriminative strength and interdependency of gait features,method accuracies,gait feature databases and likelihood ratio estimations;(2)to compare automated and observer-based gait recognition methods;to design(3)an international standard method with known validit
基金This study was supported by the General Program of National Natural Science Foundation of China[grant number 81625013 and 81772028]the Shanghai Outstanding Academic Leaders Plan[grant number 2017485]the Shanghai Talent Development Funding[grant number 2017115].
文摘The Investigator 24plex QS Kit(QIAGEN,Hilden,Germany)is a 6-dye fluorescent chemistry short tandem repeat(STR)polymerase chain reaction(PCR)amplification system that simultaneously amplifies 20 of the expanded Combined DNA Index System(CODIS)core STR loci,SE33,DYS391,and the standard sex-determining locus,amelogenin,as well as two special internal performance quality sensor controls(QS1 and QS2),which are included in the primer mix to check the PCR performance.This study was designed to be a pilot evaluation of this STR-PCR kit in a Chinese Han population regarding the PCR conditions,sensitivity,precision,accuracy,repeatability,reproducibility,and concordance;tolerance to PCR inhibitors;applicability to real“forensic-type”samples;species specificity;mixture,balance and stutter analyses,and utility in a population investigation.The exhaustive validation studies demonstrated that the Investigator 24plex QS system is accurate,sensitive and robust for STR genotyping.In addition,these genetic markers in the population data in our study indicated that they can also be useful for forensic identification and paternity testing in the Chinese Han population.
文摘目的对构建的38重InDel快速族群推断体系进行优化,根据DNA分析方法科学工作组(Scien⁃tific Working Group on DNA Analysis Method,SWGDAM)指南进行性能验证,并对其应用于东亚、欧洲、非洲及其混合人群族群推断的准确性进行验证。方法以DNA标准品9947A为模板,通过调整引物平衡性、Mg2+终浓度、优化PCR热循环参数和扩增体积等建立最优扩增条件,比较样本的等位基因丢失、非特异性扩增以及推断样本来源是否与已知信息匹配,评价该体系的相关性能。结果本体系的最佳模板用量为0.125~2 ng DNA,InDel分型结果准确,扩增均衡性好,种属特异性好;对混有血红蛋白(≤80μmol/L)、靛蓝(≤40 mmol/L)、钙离子(≤1.0 mmol/L)以及腐殖酸(≤90 ng/μL)等抑制剂的样本具有一定耐受性;可以直接扩增检测血卡、唾液卡,且族群推断结果准确;能够区分两样本的混合DNA样本;对实际案例常见生物检材的检验结果良好。结论38重InDel快速族群推断体系分型结果准确可靠,体系性能符合SWGDAM指南要求,可以准确推断未知个体的非洲、欧洲、东亚及欧亚混合人群来源,可用于法医鉴定实践。
文摘This article describes a newly devised autosomal short tandem repeat(STR)multiplex polymer-ase chain reaction(PCR)system for 19 autosomal loci(D12S391,D13S317,D16S539,D18S51,D19S433,D2S1338,D21S11,D3S1358,D5S818,D6S1043,D7S820,D8S1179,CSF1PO,FGA,TH01,TPOX,vWA,Penta D and Penta E),27 Y-chromosome STR loci(DYS19,DYS385,DYS3891,DYS38911,DYS390,DYS391,DYS392,DYS393,DYS437,DYS438,DYS439,DYS448,DYS449,DYS456,DYS458,DYS460,DYS481,DYS518,DYS533,DYS570,DYS576,DYS635,DYS627,YGATAH4 and DYF387S1)and amelogenin with six-colour fluorescent labelling.Various parameters were evaluated,such as its accuracy,sensitivity,specificity,stability,ability to ana-lysis of mixtures and effects of changes in the PCR-based procedures.All of the 47 selected STR loci were accurately and robustly amplified from 282 bloodstain samples.The species-spe-cificity was high and some ability to inhibit Hematin was identified.The lowest detectable DNA amount was ≥0.125 ng.All of the male loci of the secondary component were revealed precisely when the control DNA was mixed at male/female and male/male ratios of 1:4 or more.We conclude that the present 19-plex autosomal STR and 27 Y-STR assay is both accur-ate and sensitive.It constitutes an additional powerful tool for forensic applications.
文摘The EX 16+22Y system is a polymerase chain reaction(PCR)-based amplification kit that enables typing of 15 autosomal short tandem repeat(STR)loci(i.e„D3S1358,D13S317,D7S820,D16S539,TPOX,TH01,D2S133&CSF1PO,D19S433,vWA,D18S51,D21S11,D8S1179,D5S81&and FGA)and 22 widely used Y chromosome STR(Y-STR)loci(DYS391,DYS527a/b,DYS635,DYS458,DYS456,DYS385a/b,DYS43&DYS44&DYS437,DYS19,DYS576,DYS533,DYS393,DYS389I/n,DYS439,DYS392,Y_GATA_H4,DYS390,and DYS481)which contains 20 core Y-STR recommended by the Ministry of Public Security and amelogenin.This multiplex system was designed for the simultaneous analysis of amelogenin-Y allele mutation,single-source searches,kinship(including familial searching),mixture profiles,international data sharing,and other forensic applications.In this study,the multiplex system was validated for sensitivity of detection,species specificity,DNA mixtures,stability,sizing precision,stutter,reproducibility,and PCR-based conditions according to the Scientific Working Group on DNA Analysis Methods developmental validation guidelines and Chinese criteria for the human fluorescent STR multiplex PCR reagent.The results show that the EX16+22Y system is a robust and reliable amplification kit which can be used for human identification testing.
