单个细胞DNA样本的制备及其在灵敏度分析中的应用研究

目的建立一种用于高精确灵敏度分析的微量DNA样本制备方法。方法用显微捕获技术制备以细胞为单位的DNA样本,并用于ProfilerPlus试剂盒及ABI310遗传分析仪的灵敏度测试。结果建立了单细胞捕获操作方案,成功制备了依次含1～11个细胞的一组DNA分析样本,并可准确用于ProfilerPlus誖试剂盒及ABI310遗传分析仪的灵敏度研究。结论基于显微操作技术的高精确微量DNA分析样本制备方法可准确用于分析技术方法的灵敏度研究     

DNA IQ^TM系统在混合斑精子DNA检验中的应用

笔者在强奸案件混合斑精子的DNA检验中,以差异裂解法第一步消化去除女性细胞得到精子沉淀后,采用DNA IQTM系统直接提取混合斑中精子DNA,在200余份检材的检验中获得较满意结果。现将方法报告如下:1材料与方法1.1材料取自本实验室日常检案积累的混合斑样本216份。主要仪器和试剂采用9700PCR扩增仪(美国ABI公司)、ABI3100型遗传分析仪(美国ABI公司)、DNA IQTM系统(Promega公司)、AmpF1STRIdentifiler试剂盒((美国ABI公司)。1.2方法1.2.1 DNA提取参照差异裂解法,经过套管离心去除女性细胞后,得到沉淀精子,加入100~200ulDNA …     

Identifiler体系单基因排除的三联体亲权1例

1案例王某,男,33岁,1999年与前妻张某离婚,婚生子判由王某抚养。后王某再婚,但三年来一直没有生育,去医院检查得知自己生育能力极低,遂怀疑前妻所生之子不是自己亲生,来我处申请进行亲子鉴定。2检验分别抽取王某、张某与其子的静脉血各0.1mL,EDTA抗凝。用Chelex-100法提取上述血样的DNA,用ABI公司IdentifilerKit荧光标记复合扩增试剂盒扩增,ABI310型基因分析仪对扩增产物进行检测,Genotyperv2.5.2软件进行基因分型,所得STR基因型结果见表1。由表1可见在所检测的15个STR基因座中仅有D16S539位点孩子与王某不符合遗传关系。遂采用…     

用Chelex100法及有机溶剂提取法联合提取DNA扩增STR基因座的比较研究

为了比较不同方法提取DNA扩增STR基因座的成功率 ,本文用Chelex10 0法及Chelex10 0法联合有机溶剂提取法分别提取性犯罪案件 6份血斑及 6份混合斑中精子DNA ,并用PE公司ProfilerPlus系统复合扩增 ,ABI 310型基因分型仪检测。结果表明 ,常规采用Chelex10 0法提取血斑及精斑等生物检材DNA作STR基因座检验失败或所检测的位点较少时 ,应对Chelex10 0法提取的DNA上清液再用有机溶剂提取法提取 ,可极大提高DNA检验成功率。采用本文建立的Chelex10 0法联合有机溶剂提取法 ,提高了PCR扩增阳性率 ,对实际检案很有帮助 ;在PCR扩增前应常规进行DNA定量检验 ,否则对于重要检材应进行梯度扩增。     

wizard clean up在混合斑DNA检验中的应用

目的建立一种快速、简单、有效的混合斑DNA检验方法。方法在100例检案中,利用wizardcleanup直接对精子消化液进行纯化浓缩。采用profilerplus试剂盒进行复合扩增,产物经ABI310基因分析仪检测。结果从混合斑中成功获得精斑10个STR位点的DNA分型。结论wizardcleanup处理混合斑,能有效去除女性成份,得到精斑DNA进行分型。     

DNA-STR分型检验人体脱落上皮细胞2例

作者结合 2个典型案例 ,对一些特殊载体上的脱落上皮细胞成功地进行了STR检验分型 ,并就检验中的相关问题进行讨论 ,旨在进一步提高办案人员对此类检材应用价值的注意。1　案例资料案例 1　 2 0 0 3年 1月在某旅社内 ,犯罪嫌疑人王某将徐某 (女 ,2 7岁 )掐颈致死后 ,用打火机烧灼电视机电线并用手拉成多节 ,用于捆绑死者。用纱线两步擦试法转移电视机电线上上皮细胞 ,用Chelex 10 0法提取上皮细胞DNA ,Microcon 10 0纯化柱处理 ,用AmpFISTR profilerPlusTM 系统复合扩增试剂盒进行STR位点的复合扩增 ,扩增产物经ABI 310型基因分析…     

wiazrd clean up在混合斑DNA检验中的应用

目的 建立一种快速、简单、有效的混合斑DNA检验方法。方法 在100例检案中，利用wizard clean up直接对于精子消化液进行纯化浓缩。采用profiler plus试剂盒进行复合扩增，产物经ABI310基因分析仪检测。结果 从混合斑中成功获得精斑10个STR位点的DNA分型。结论 wizard clean up处理混合斑，能有效去除女性成份，得到精斑DNA进行分型。     

3种STR试剂盒在二联体亲缘鉴定中联合应用1例

1案例资料 1．1样本及检验方法 要求进行亲缘关系鉴定的王某（男,42岁）和李某（女,13岁）的血液样本各1份。采用Chelex．100法提取模板DNA。分别采用Identifiler试剂盒（ABI公司,美国）、PowerPlex 21试剂盒（Promega公司,美国）、STRtyper-10G试剂盒（珠海科登生物工程有限公司）进行分型检验。     

人体死后体内氰菊酯及其代谢产物分布检测1例

1案例资料1.1简要案情池某,女。2011年11月6日死亡,因死因不明怀疑被人投毒。提取心血未检出毒物成分,后解剖提取肾、心、子宫、肝、脑、肺等脏器,要求进行毒物分析检验。1.2毒物分析检验主要仪器与试剂美国ABI公司API2000 LCMS/MS;岛津LC-20A型高效液相色谱仪;甲氰菊酯、3-苯氧基苯甲酸（3-phenoxybenzoic acid,3-PBA）和甲     

微量生物检材DNA检验的研究

在日常检案中经常遇到微量的生物检材 ,由于重视不够 ,提取不当 ,检验效果不甚理想。为了充分发挥微量生物检材的作用 ,更好的为侦察破案服务 ,笔者对部分微量生物检材进行了PCR STR检验研究 ,并取得了一定效果。现报道如下 :1　方法和结果实验样品　均由非本实验室的志愿者提供 ,一次性牙刷 10个 ,日常用牙刷 4个 ,头皮屑 2人分 ,一次性纸杯 10个 ,面部擦拭斑迹 5份 ,梳子 2把。另取杀人案件现场嫌疑人遗留的衣服领子 1个 ,盗窃案件现场提取的犯罪嫌疑人饮酒用的瓷碗 1个。试剂与仪器采用 970 0扩增仪 (ABI公司 )及ABI 377测序仪 ,Pr…     

Strengthening the Connection Between Acquired Brain Injury (ABI) and Family Violence: The Importance of Ongoing Monitoring,Research and Inclusive Terminology

Family violence (FV) harms communities worldwide so FV prevention strategies and effective responses are urgently needed. This article reports on FV apparent in a study which explored the experiences of people with both ABI and justice system encounters in Victoria, Australia. One hundred interviews and one focus group consulted people with ABI, their families and carers, and various stakeholder groups in the Victorian justice system in Australia. Qualitative content analysis determined dominant themes and sub-themes and the less common themes. Inductive interpretive content analysis identified themes commonly found in previous published research and themes that appeared unique to, or unanticipated in, our data, such as the FV theme upon which this article focuses. Our findings reveal that FV has adversely affected many people with ABI who came into contact with Victoria’s justice system. Further, as ABI and FV often co-occur with substance abuse, mental health problems, socio-economic and many other significant disadvantages, for some FV perpetrators with an ABI, their ABI symptoms and characteristic co-morbidities may be a mitigating factor in their offending. The connection between ABI and family violence emerged as a troubling research theme. Indeed, the impact of FV on too many of our participants with an ABI compels us to call for further related research and secondary prevention programs targeted at FV victims, and offenders, living with ABI. An intersectional understanding of family violence and TBI/ABI in social ecological contexts is required to better understand brain injury at both individual and population levels.

     

Evaluation of a multicapillary electrophoresis instrument for mitochondrial DNA typing

Laser-induced detection of fluorescent labeled PCR products and multi-wavelength detection (i.e., multicolor analysis) enables rapid generation of mtDNA sequencing profiles. Traditionally, polyacrylamide slab gels have been used as the electrophoretic medium for mtDNA sequencing in forensic analyses. Replacement of slab gel electrophoresis with capillary electrophoresis (CE) can facilitate automation of the analytical process. Automation and high throughput can be further enhanced by using multicapillary electrophoretic systems. The use of the ABI Prism 3100 Genetic Analyzer (ABI 3100, Applied Biosystems, Foster City, CA) as well as the ABI Prism 310 Genetic Analyzer (ABI 310, Applied Biosystems, Foster City, CA) were evaluated for mtDNA sequencing capabilities and compared with sequencing results obtained on the platform currently in use in the FBI Laboratory (the ABI Prism 377 DNA Sequencer, ABI 377, Applied Biosystems, Foster City, CA). Various studies were performed to assess the utility of the ABI 3100, as well as the ABI 310 for mtDNA sequencing. The tests included: comparisons of results obtained among the ABI 3100, the ABI 310 and the ABI 377 instruments; comparisons of results obtained within and between capillary arrays; evaluation of capillary length; evaluation of sample injection time; evaluation of the resolution of mixtures/heteroplasmic samples; and evaluation of the sensitivity of detection of a minor component with reduced template on the ABI 3100. In addition, other studies were performed to improve sample preparation; these included: comparison of template suppression reagent (TSR, Applied Biosystems, Foster City, CA) versus formamide; the use of Performa DTR Gel Filtration Cartridges (Edge BioSystems Inc., Gaithersburg, MD) versus Centri-Sep Spin Columns (Princeton Separations, Adelphia, NJ) for product purification after cycle sequencing; and sample stability after denaturation. The data support that valid and reliable results can be obtained using either capillary electrophoresis instrument, and the quality of sequencing results are comparable to or better than those obtained from the ABI 377 instrument.     

Comparison of DNA typing using AmpFlSTR Yfiler and PowerPlex Y System,for specimens subject to very long storage

We performed DNA typing from blood stains stored at room temperature for 22–30 years, using AmpFlSTR Yfiler PCR amplification Kit and PowerPlex Y System to extract and amplify the DNA and performing electrophoresis with an ABI 310 Genetic Analyzer. We identified alleles using GeneMapper ID v3.2.1 software. We found that long-term storage tended to reduce the number of detectable loci in the blood stains and that it was easier to detect the loci of smaller amplicons than larger amplicons.     

PCR扩增体系的体积减少对DNA分析的影响

目的探讨PCR扩增体系的体积减少对DNA分析的影响。方法4份样品采用ProfilerPlus试剂盒,在同样条件下,对50μl、25μl、12.5μl、6.25μl四种体积的体系进行扩增,扩增产物分别经ABIPRISMTM310型基因分析仪、ABIPRISMTM377型测序仪、ABIPRISMTM3100型基因分析仪电泳,并经GeneScan3.1基因扫描软件和Genotyper2.5分析软件分析得到实验结果。结果小体积的扩增体系容易出现等位基因的丢失、额外等位基因的产生等现象。结论在检材情况较差时,应该慎用小体积的扩增体系。     

混合生物样品的组分分析及其STR基因型判定

目的应用荧光标记STR多基因座联合检测技术对混合生物样品较少组分最低比例检出限和STR基因型的判定进行研究。方法 将已知浓度的人标准细胞系DNA:9947A和K562分别按照1:1、1:4、1:9、1:19、1:39、1:59、1:79和1:99比例混合后作为扩增模板,应用Profiler plus试剂盒对D3S1358、VWA、FGA、D8S1179、D21S11、D18S51、D5S818、D13S317、D7S820和性别鉴定基因座检验。结果在比例为1:19时能明确判定两个体各基因座基因型,且两样品基因型结果峰高平均值之比与样品浓度之比正相关。结论应用荧光标记STR多基因座联合检测技术可以对一定比例的混合生物样品进行STR基因型判定,并对其混合比例状况进行大致推断。     

Abuse behavior inventory: cutpoint, validity, and characterization of discrepancies

This study determines a clinical cutting score for the 29-item Abuse Behavior Inventory (ABI) developed by Shepard and Campbell (1992) to measure both physical and psychological abuse experiences. The authors report on a sample of 392 White and African American women from primary care waiting rooms, who completed the ABI and the revised Conflict Tactics Scale (CTS2). An ABI cutoff score of 10 maximizes validity and produces a sensitivity of 77% and a specificity of 81% compared to the CTS2. Cronbach's alpha is .92 for the entire scale, .91 for the psychological subscale, and .86 for the physical subscale. Correlation between the ABI and CTS2 is .76, with subscale correlations of .74 between ABI psychological and CTS2 verbal aggression, and .71 between ABI physical and CTS2 physical aggression, injury, and sexual coercion.     

国产NuHi-POP4与ABI POP-4在毛细管电泳分离效果的比较

目的比较国产基因分离胶NuHi-POP4与美国产商品ABI POP-4在法医鉴定应用中的分离性能。方法以国产基因分离胶NuHi-POP4和美国产的ABI POP-4为研究对象,分别考察其在毛细管电泳过程中对标准品LIZ500和实际样品的分离效果,并统计其电泳参数,包括电泳时间、电流、单碱基分辨率以及线性关系等,从而来比较两种胶的分离性能,评价其性能。结果 NuHi-POP4与ABI POP-4相比,不仅可以显著缩短内标和样品的分析时间,同时还表现出电流稳定、分辨率高、线性关系好等特点。结论国产NuHi-POP4与美国ABI POP-4相比,同样具有稳定、快速、高分辨等优势,结果准确可靠。     

Allele frequencies of dinucleotide repeat marker loci on the X chromosome in the Japanese population

The allele frequencies of 18 dinucleotide repeat marker loci on the X chromosome have been analyzed in 130 Japanese individuals living in Kanagawa by means of multiplex PCR and the ABI PRISM Linkage Mapping Set MD 10 Panel 28, followed by capillary electrophoresis using the ABI PRISM 310 Genetic Analyzer.     

Physical qualification of drivers; medical examination; certificate. Federal Motor Carrier Safety Administration (FMCSA), DOT. Final rule

This document updates and simplifies the medical examination form that is currently used to determine the physical qualification of commercial motor vehicle (CMV) drivers operating in interstate commerce. The FMCSA takes this action in response to numerous requests from medical examiners to update and simplify the medical examination form that is currently used. This action is intended to reduce the incidence of errors on such forms and to provide more uniform medical examinations of CMV drivers engaged in interstate commerce. The current Federal physical qualification standards tested by medical examiners and recorded on the form will not be revised in this rulemaking.