The DNATyper^(TM)21 kit has good species specificity,typing accuracy,and locus amplification balance,which can be used for forensic genetic analysis such as individual identification and paternity testing.To improve t...The DNATyper^(TM)21 kit has good species specificity,typing accuracy,and locus amplification balance,which can be used for forensic genetic analysis such as individual identification and paternity testing.To improve the anti-inhibitor ability of this kit,the addition of bovine serum albumin(BSA),bovine thrombin(BT),FastStart Taq DNA Polymerase,TaKaRa Ex Taq Hot Start Version(Ex Taq HS),MyFiTM DNA Polymerase,and Klentaq DNA Polymerase(Klentaq)as PCR enhancers to the PCR reaction system was explored in the presence of different concentrations of inhibitors,such as indigo,humic acid(HA),hemoglobin,heme,and melanin.The results revealed that BSA exhibited high efficiency in improving the detection rate of STR loci;BT only helped to overcome the typing inhibition caused by indigo,melanin,HA,and heme;Four types of DNA polymerase had different anti-inhibitor effects on different inhibitors,among which Ex Taq HS was more effective than the other three DNA polymerases,but displayed comparatively lower resistance to hemoglobin than BSA and BT.This study provides basic data for further optimization of the kit through comprehensive analysis and facilitates its application in daily forensic investigations.展开更多
We describe the optimization and validation of the DNATyper^(TM)15 multiplex polymerase chain reaction(PCR)genotyping system for autosomal short tandem repeat(STR)amplification at 14 autosomal loci(D6S1043,D21S11,D7S8...We describe the optimization and validation of the DNATyper^(TM)15 multiplex polymerase chain reaction(PCR)genotyping system for autosomal short tandem repeat(STR)amplification at 14 autosomal loci(D6S1043,D21S11,D7S820,CSF1PO,D2S1338,D3S1358,D13S317,D8S1179,D16S539,Penta E,D5S818,vWA,D18S51,and FGA)and amelogenin,a sex‑determining locus.Several DNATyper^(TM)15 assay variables were optimized,including hot start Taq polymerase concentration,Taq polymerase activation time,magnesium concentration,primer concentration,annealing temperature,reaction volume,and cycle number.The performance of the assay was validated with respect to species specificity,sensitivity to template concentration,stability,accuracy,influence of the DNA extraction methods,and the ability to genotype the mixture samples.The performance of the DNATyper^(TM)15 system on casework samples was compared with that of two widely used STR amplification kits,Identifiler^(TM)(Applied Biosystems,Carlsbad,CA,USA)and PowerPlex 16®(Promega,Madison,WI,USA).The conditions for PCR‑based DNATyper^(TM)15 genotyping were optimized.Contamination from forensically relevant nonhuman DNA was not found to impact genotyping results,and full profiles were generated for all the reactions containing≥0.125 ng of DNA template.No significant difference in performance was observed even after the DNATyper^(TM)15 assay components were subjected to 20 freeze‑thaw cycles.The performances of DNATyper^(TM)15,Identifiler^(TM),and PowerPlex 16®were comparable in terms of sensitivity and the ability to genotype the mixed samples and case‑type samples,with the assays giving the same genotyping results for all the shared loci.The DNA extraction methods did not affect the performance of any of the systems.Our results demonstrate that the DNATyper^(TM)15 system is suitable for genotyping in both forensic DNA database work and case‑type samples.展开更多
目的研究福建三明汉族人群20个常染色体STR基因座遗传多态性,评估其法医学中的应用价值。方法采用Microreader^(TM)21 Direct ID System试剂盒对三明汉族508例无关个体血样进行检测,获取20个基因座的数据资料,分析基因分布频率及群体遗...目的研究福建三明汉族人群20个常染色体STR基因座遗传多态性,评估其法医学中的应用价值。方法采用Microreader^(TM)21 Direct ID System试剂盒对三明汉族508例无关个体血样进行检测,获取20个基因座的数据资料,分析基因分布频率及群体遗传学参数。结果20个STR基因座的等位基因频率为(0.0010~0.5827),各基因座基因型分布符合Hardy-Weinberg平衡(P>0.05);多肽信息总量(PIC)为0.52~0.91;期望杂合度(He)为0.5965~0.9173;匹配率(PM)为0.0313~0.2339;个体识别能力(DP)为0.766~0.985;排除概率(PE)为0.287~0.831;累计个体识别能力(TDP)为1-1.1124×10^(-24)=0.999999999999999999999999;累计排除率(CPE)为1-2.4513×10^(-9)=0.999999998。结论20个常染色体STR基因座在三明汉族人群中呈高度多态性和较好的鉴别能力,对个体识别及亲权鉴定具有较大法医学应用价值。展开更多
文摘The DNATyper^(TM)21 kit has good species specificity,typing accuracy,and locus amplification balance,which can be used for forensic genetic analysis such as individual identification and paternity testing.To improve the anti-inhibitor ability of this kit,the addition of bovine serum albumin(BSA),bovine thrombin(BT),FastStart Taq DNA Polymerase,TaKaRa Ex Taq Hot Start Version(Ex Taq HS),MyFiTM DNA Polymerase,and Klentaq DNA Polymerase(Klentaq)as PCR enhancers to the PCR reaction system was explored in the presence of different concentrations of inhibitors,such as indigo,humic acid(HA),hemoglobin,heme,and melanin.The results revealed that BSA exhibited high efficiency in improving the detection rate of STR loci;BT only helped to overcome the typing inhibition caused by indigo,melanin,HA,and heme;Four types of DNA polymerase had different anti-inhibitor effects on different inhibitors,among which Ex Taq HS was more effective than the other three DNA polymerases,but displayed comparatively lower resistance to hemoglobin than BSA and BT.This study provides basic data for further optimization of the kit through comprehensive analysis and facilitates its application in daily forensic investigations.
基金the National Science and Technology Supporting Program during the 10th 5‑year plan period(2001BA801B02)。
文摘We describe the optimization and validation of the DNATyper^(TM)15 multiplex polymerase chain reaction(PCR)genotyping system for autosomal short tandem repeat(STR)amplification at 14 autosomal loci(D6S1043,D21S11,D7S820,CSF1PO,D2S1338,D3S1358,D13S317,D8S1179,D16S539,Penta E,D5S818,vWA,D18S51,and FGA)and amelogenin,a sex‑determining locus.Several DNATyper^(TM)15 assay variables were optimized,including hot start Taq polymerase concentration,Taq polymerase activation time,magnesium concentration,primer concentration,annealing temperature,reaction volume,and cycle number.The performance of the assay was validated with respect to species specificity,sensitivity to template concentration,stability,accuracy,influence of the DNA extraction methods,and the ability to genotype the mixture samples.The performance of the DNATyper^(TM)15 system on casework samples was compared with that of two widely used STR amplification kits,Identifiler^(TM)(Applied Biosystems,Carlsbad,CA,USA)and PowerPlex 16®(Promega,Madison,WI,USA).The conditions for PCR‑based DNATyper^(TM)15 genotyping were optimized.Contamination from forensically relevant nonhuman DNA was not found to impact genotyping results,and full profiles were generated for all the reactions containing≥0.125 ng of DNA template.No significant difference in performance was observed even after the DNATyper^(TM)15 assay components were subjected to 20 freeze‑thaw cycles.The performances of DNATyper^(TM)15,Identifiler^(TM),and PowerPlex 16®were comparable in terms of sensitivity and the ability to genotype the mixed samples and case‑type samples,with the assays giving the same genotyping results for all the shared loci.The DNA extraction methods did not affect the performance of any of the systems.Our results demonstrate that the DNATyper^(TM)15 system is suitable for genotyping in both forensic DNA database work and case‑type samples.
文摘目的研究福建三明汉族人群20个常染色体STR基因座遗传多态性,评估其法医学中的应用价值。方法采用Microreader^(TM)21 Direct ID System试剂盒对三明汉族508例无关个体血样进行检测,获取20个基因座的数据资料,分析基因分布频率及群体遗传学参数。结果20个STR基因座的等位基因频率为(0.0010~0.5827),各基因座基因型分布符合Hardy-Weinberg平衡(P>0.05);多肽信息总量(PIC)为0.52~0.91;期望杂合度(He)为0.5965~0.9173;匹配率(PM)为0.0313~0.2339;个体识别能力(DP)为0.766~0.985;排除概率(PE)为0.287~0.831;累计个体识别能力(TDP)为1-1.1124×10^(-24)=0.999999999999999999999999;累计排除率(CPE)为1-2.4513×10^(-9)=0.999999998。结论20个常染色体STR基因座在三明汉族人群中呈高度多态性和较好的鉴别能力,对个体识别及亲权鉴定具有较大法医学应用价值。
