Jovanovich Stevan, Bogdan Greg, Belcinski Richard, Buscaino Jacklyn, Burgi Dean, Butts Erica L R, Chear Kaiwan, Ciopyk Brian, Eberhart David, El-Sissi Omar, Franklin Helen, Gangano Stefanie, Gass Jennifer, Harris Dennis, Hennessy Lori, Kindwall Alex, King David, Klevenberg Jim, Li Yuan, Mehendale Neelima, McIntosh Roger, Nielsen Bill, Park Charles, Pearson Francesca, Schueren Robert, Stainton Nancy, Troup Charles, Vallone Peter M, Vangbo Mattias, Woudenberg Timothy, Wyrick David, Williams Stephen
IntegenX Inc. 5720 Stoneridge Drive, Suite 300, Pleasanton, CA 94588-2739, USA.
IntegenX Inc. 5720 Stoneridge Drive, Suite 300, Pleasanton, CA 94588-2739, USA.
Forensic Sci Int Genet. 2015 May;16:181-194. doi: 10.1016/j.fsigen.2014.12.004. Epub 2015 Jan 8.
Short tandem repeat (STR) DNA typing is a global standard for human identification. Current practice involves highly trained forensic analysts, operating in a laboratory setting, using multiple instruments to process samples and analyze the data. Here, we report the developmental validation of a fully integrated and automated DNA profiling system, the RapidHIT® System, capable of producing up to five high quality STR profiles with full controls in approximately 90min using PowerPlex®16 HS RapidHIT chemistry. The system integrates all sample handling steps: starting from lysis of cells on buccal swabs or other buccal sample types through DNA extraction, normalization, amplification,capillary array electrophoresis, detection, and integrated software analysis. The results describe the developmental validation of the RapidHIT™ System for buccal samples processed with the DNA IQ™ extraction chemistry using a guandinium chaotropic agent and paramagnetic beads followed by amplification using a modified version of PowerPlex 16 HS chemistry (PowerPlex 16 HS RapidHIT chemistry), and capillary electrophoresis with manual review of genotyping data following interpretation guidelines. All processing from the buccal swab to generation and processing of the profile occurs on the RapidHIT platform.
are concordant with traditional methods, with 88% first pass success rates for both the CODIS and PowerPlex 16 loci. Average peak height ratios were 0.89 for buccal swabs. The system produces full profiles from swabs with at least 176 ng of saliva DNA. Rapid DNA identification systems will significantly enhance capabilities for forensic labs, intelligence, defense, law enforcement, refugee and immigration applications, and kinship analysis.
短串联重复序列(STR)DNA分型是人类身份识别的全球标准。目前的做法需要训练有素的法医分析师在实验室环境中操作,使用多种仪器处理样本并分析数据。在此,我们报告了一种完全集成且自动化的DNA分析系统RapidHIT®系统的开发验证情况,该系统使用PowerPlex®16 HS RapidHIT化学方法,能够在约90分钟内生成多达五个带有完全对照的高质量STR图谱。该系统集成了所有样本处理步骤:从口腔拭子或其他口腔样本类型的细胞裂解开始,经过DNA提取、标准化、扩增、毛细管阵列电泳、检测以及集成软件分析。结果描述了使用含有胍盐变性剂和顺磁珠的DNA IQ™提取化学方法处理口腔样本,随后使用改良版PowerPlex 16 HS化学方法(PowerPlex 16 HS RapidHIT化学方法)进行扩增,并按照解释指南对基因分型数据进行人工审核后进行毛细管电泳,以此对RapidHIT™系统进行开发验证。从口腔拭子到图谱生成和处理的所有过程均在RapidHIT平台上进行。
与传统方法一致,联合DNA索引系统(CODIS)和PowerPlex 16位点的首次通过率均为88%。口腔拭子的平均峰高比为0.89。该系统能够从含有至少176 ng唾液DNA的拭子中生成完整图谱。快速DNA识别系统将显著提高法医实验室、情报、国防、执法、难民和移民应用以及亲缘关系分析的能力。