法庭科学SWGDAM验证流程课件

Click to Edit Master Title,Click to edit Master text,Second level,Third level,Fourth level,Fifth level,#,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,2019/1/2,#,法庭科学,SWGDAM,验证流程,法庭科学SWGDAM验证流程,议程,SWGDAM,介绍,华夏,TM,白金,SWGDAM,验证,YfilerPlus,TM,SWGDAM,验证,ISO,18385,介绍,议程SWGDAM 介绍,SWGDAM,主页,Source: swgdam.org,SWGDAM主页Source: swgdam.org,关于,SWGDAM,First,meeting,of Technical,Working Group on DNA Analysis Methods (TWGDAM,) was held in,November,1988, with a subcommittee on,restriction fragment length polymorphism (RFLP) DNA analysis.,TWGDAM continued to provide a level of direction to the forensic DNA community by issuing guidelines for DNA analysis, and the guidelines became de facto standards and were recognized by courts.,SWGDAM Chairman is appointed by the FBI Director, and members are,selected by,the Chairman,from,forensic DNA laboratories,.,SWGDAM has multiple committees including Autosomal STR Committee, CODIS Committee, Rapid DNA Committee, Next Generation Sequencing Working Group, etc.,One of SWGDAMs most important responsibilities is the recommendation of revisions to the FBIs Quality Assurance Standards (QAS) for DNA Analysis,.,关于SWGDAMFirst meeting of Techn,SWGDAM,出版物,SWGDAM 出版物,SWGDAM,法医,DNA,分析方法验证指,导,3.7 Population Studies,3.8 Mixture Studies,3.9 PCR Based Studies,3.3 Sensitivity,3.4 Stability,3.5.,Precision,& Accuracy,3.6 Case Type Samples,3.2 Species Specificity,3.1 Genetic Markers,SWGDAM Validation Guidelines for Forensic DNA Analysis Methods approved in November 2012,www.swgdam.org,SWGDAM法医DNA分析方法验证指导3.7 Populat,华夏白金物种特异性,非灵长类,10 ng non-primate DNA, or 105 copies of microbes (species,commonly found in the,oral cavity, listed in User Guide),were added to the system using 29 PCR cycles.,Low peaks were detected with bovine, dog (like contamination, also found in the repeat), horse, mouse and pig.,Bovine,Chicken,Dog,Horse,Microbes,Mouse,Pig,Rabbit,Rat,Sheep,NTC,200 rfu,200 rfu,700 rfu,SWGDAM 3.2,The,ability to detect genetic information from non-targeted species (e.g., detection of microbial DNA in a human assay) should be,determined.,华夏白金物种特异性 非灵长类10 ng non-prim,华夏白金一次性成功通过率,All sample types,have 90%,pass rate (PAT = 175 rfu) with 1 Proportion test.,The recommended rxn volume for HXP is 25ul, some tests,here were,done with 10ul reaction volume to challenge the system.,SWGDAM 3.4,The ability to obtain results from DNA recovered from biological samples deposited on various substrates and subjected to various environmental and chemical insults should be evaluated.,华夏白金一次性成功通过率All sample types h,样本准在,3500xl,上的准确度,Size deviation of 42 population samples showed deviation 0.50bp for all alleles,SWGDAM 3.5,Precision and accuracy,of the assay should be,demonstrated.,Accuracy,is the degree of conformity of a measured quantity to its actual (true) value,.,样本准在3500xl上的准确度Size deviation,遗,传标记在不同种族的人群中的分布,SWGDAM 3.7,The distribution of genetic markers in populations should be determined in relevant population groups.,Allele frequency for major Chinese population groups was studied.,Data of D6S1043 was presented above, and the data with other markers was published in UG.,遗传标记在不同种族的人群中的分布 SWGDAM 3.7 Th,反应预混物保护带研究,Eight raw materials in master mix were varied up to 20%.,ICB was,above 35%,for most raw materials with,10% or even 20% variation,.,Detail data including capability analysis,presented,next.,Component concentrations,Individual components,The Huaxia Platinum MM is robust to raw material change,SWGDAM 3.9,The reaction conditions needed to provide the required degree of specificity and robustness should be determined.,反应预混物保护带研究Eight raw materials,SWGDAM,对,Y-STR,的指导,Y-STR typing is also used in lieu of autosomal typing for the,detection of male DNA in mixtures that contain an overabundance of female DNA,. Given that under certain conditions a male minor contributor in a mixture of female:male DNA may only be detectable by Y-STR typing, laboratories should pursue Y-STR analysis as the most appropriate means of detecting a male contributor(s) in some forensic samples,Interpretation Guidelines for Y-Chromosome STR Typing approved in January 2014,www.swgdam.org,This SWGDAM statement highlights the importance of male specificity of Y-STR kit with the male female mixtures commonly found with the sex assault cases.,SWGDAM 对Y-STR的指导Y-STR typing i,Primer mix,Design/screen primer pairs with minimal female cross-reactivity,Master mix,Lower the Mg and Taq enzyme concentrations to minimize female cross-reactivity,Multiple rounds of DOE were performed to optimize the MM components,PCR condition,Increase the annealing/extension temperature to increase male specificity,提高,Y-,特异性的工作,Primer mix提高Y-特异性的工作,Yfiler Plus,在男女混合,DNA,的表现,Yfiler Plus recovered full profile with 1:1000 male:female mixture,No extra peaks with Yfiler Plus, while multiple extra peaks with PPY23 and Yfiler,Internal Product Spec: The kit will recover full profile with average 40% ICB when amplified with male:female mixtures of 1:1000,Yfiler Plus 在男女混合DNA的表现Yfiler,Yfiler Plus,高浓度女性,DNA,下的基线,200rfu,Low female cross-reactivity (175 PAT) within the read region are all documented in the UG.,Blue 271,Red 139/144,Red grass,Outside the read region,68 bp,406 bp,Internal Product Spec:,The Yfiler Plus kit shall not amplify reproducible female DNA in the 1 ug range, without reproducible drop ins & artifacts within the read region.,Yfiler Plus高浓度女性DNA下的基线200rfuL,3,微克女性,DNA,在三个盒子的比较,3 ug of DNA from 10 different female donors were tested with the Yfiler Plus, PPY23 and Yfiler kits. The highest reproducible female artifacts from each sample were plotted by kit. Yfiler Plus and Yfiler have significantly lower artifact peak heights than PPY23.,PAT,3 微克女性DNA在三个盒子的比较3 ug of DNA f,新的法医,DNA,标准,ISO 18385,Meet ISO 18385 requirements,for the manufacture of Forensic,DNA Grade,reagents,Provide same,level of manufacturing quality and QC testing across,HID,product,lines,Both Yfiler plus and Huaxia Platinum kits are ISO 18385 compliant,Manufacturing,Quality Commitment,新的法医DNA标准 ISO 18385Meet ISO,