单核苷酸多态性(SNP)基因分型技术可以产生用于人类遗骸鉴定的调查线索,包括生物性别的估计,生物地理祖先(BGA),外部可见特性(EVC),身份,单亲血统和扩展的亲属关系。ForenSeq®KintelligenceKit为法医实验室提供了将这套遗传工具应用于法医样本的能力,使用一个针对10,230个SNP的小组(包括56个祖先信息,24表型信息,94身份信息,106X染色体,85个Y染色体和9867个亲缘信息SNP)在MiSeqFGx®测序系统上测序。ForenSeq®KintelligenceKit已经过内部验证,由澳大利亚联邦警察国家身份不明和失踪人员DNA计划(AFP计划)进行优化和操作,以进行死因调查。内部验证是根据DNA分析方法科学工作组指南(不包括混合物分析)进行的,专注于人类遗骸识别案例中通常遇到的样本类型,比如骨头,牙齿,钉,血和头发该工作流程针对高通量文库制备和测序工作流程进行了优化,并开发了其他分析阈值以提高低DNA输入样品的基因分型准确性。此外,将从试剂盒中产生的遗传智力与自我声明的生物性别进行比较,DNA供体的EVC和BGA评估一致性。该试剂盒能够产生从1.0ng到0.1ngDNA的高质量SNP谱,具有高重复性和再现性,和最小的背景噪音。生物性别的预测准确率(95%),头发颜色(58%),眼睛颜色(74%)和BGA推断(一致:74%;部分一致:10%;不确定:16%)是根据自我声明的数据确定的.此外,来自10个相关个体的志愿者家庭组的SNP概况被上传到GEDmatchPRO™以评估亲属关系的准确性。该试剂盒能够检测(97%)和准确分类(90%)从第一到第五度的遗传关系。KintelligenceKit为AFP计划提供了一个强大而可靠的遗传情报工具,用于身份不明和失踪人员的调查,它被设计为使用现有的实验室仪器在单个多重测定中对多个具有挑战性的样品进行测序。
Single nucleotide polymorphism (SNP) genotyping technologies can generate investigative leads for human remains identification, including estimation of biological sex, biogeographical ancestry (BGA), externally visible characteristics (EVCs), identity, uniparental lineage and extended kinship. The ForenSeq® Kintelligence Kit provides forensic laboratories with the ability to apply this suite of genetic tools to forensic samples using one panel targeting 10,230 SNPs (including 56 ancestry-informative, 24 phenotype-informative, 94 identity-informative, 106 X chromosome, 85 Y chromosome and 9867 kinship-informative SNPs) sequenced on the MiSeq FGx® Sequencing System. The ForenSeq® Kintelligence Kit has been internally validated, optimised and operationalised by the Australian Federal Police National DNA Program for Unidentified and Missing Persons (AFP Program) for coronial casework. The internal validation was conducted according to the Scientific Working Group on DNA Analysis Methods guidelines (excluding mixture analysis), focussing on sample types typically encountered in human remains identification casework, such as bones, teeth, nail, blood and hair. The workflow was optimised for a high throughput library preparation and sequencing workflow, and additional analytical thresholds were developed to improve genotyping accuracy for low DNA input samples. Additionally, the genetic intelligence generated from the kit was compared to the self-declared biological sex, EVCs and BGA of the DNA donors to assess concordance. The kit was able to produce high quality SNP profiles from 1.0 ng down to 0.1 ng of DNA, with high repeatability and reproducibility, and minimal background noise. The prediction accuracy for biological sex (95%), hair colour (58%), eye colour (74%) and BGA inferences (consistent: 74%; partially consistent: 10%; inconclusive: 16%) was determined based on self-declared data. Additionally, SNP profiles from a volunteer family group of ten related individuals were uploaded to GEDmatch PRO™ to assess kinship accuracy. The kit was capable of detecting (97%) and accurately classifying (90%) genetic relationships spanning from first to fifth degree. The Kintelligence Kit provides the AFP Program with a robust and reliable genetic intelligence tool for unidentified and missing persons investigations, which has been designed to sequence multiple challenging samples in a single multiplexed assay using existing laboratory instrumentation.