Han Junping, Zhuang Bin, Zou Lixin, Wang Daoyu, Jiang Li, Wei Yi-Liang, Zhao Lijian, Zhao Lei, Li Caixia
Technology Department of Chaoyang Sub-bureau, Beijing Public Security Bureau, Beijing, P. R. China.
Key Laboratory of Forensic Genetics, Beijing Engineering Research Center of Crime Scene Evidence Examination, National Engineering Laboratory for Forensic Science, Institute of Forensic Science, Beijing, P. R. China.
Electrophoresis. 2024 May;45(9-10):814-828. doi: 10.1002/elps.202300187. Epub 2024 Mar 9.
Analysis of short tandem repeats (STRs) is a global standard method for human identification. Insertion/Deletion polymorphisms (DIPs) can be used for biogeographical ancestry inference. Current DNA typing involves a trained forensic worker operating several specialized instruments in a controlled laboratory environment, which takes 6-8 h. We developed the Quick TargSeq 1.0 integrated system (hereinafter abbreviated to Quick TargSeq) for automated generation of STR and DIP profiles from buccal swab samples and blood stains. The system fully integrates the processes of DNA extraction, polymerase chain reaction (PCR) amplification, and electrophoresis separation using microfluidic biochip technology. Internal validation studies were performed using RTyper 21 or DIP 38 chip cartridges with single-source reference samples according to the Scientific Working Group for DNA Analysis Methods guidelines. These results indicated that the Quick TargSeq system can process reference samples and generate STR or DIP profiles in approximately 2 h, and the profiles were concordant with those determined using traditional STR or DIP analysis methods. Thus, reproducible and concordant DNA profiles were obtained from reference samples. Throughout the study, no lane-to-lane or run-to-run contamination was observed. The Quick TargSeq system produced full profiles from buccal swabs with at least eight swipes, dried blood spot cards with two 2-mm disks, or 10 ng of purified DNA. Potential PCR inhibitors (i.e., coffee, smoking tobacco, and chewing tobacco) did not appear to affect the amplification reactions of the instrument. The overall success rate and concordance rate of 153 samples were 94.12% and 93.44%, respectively, which is comparable to other commercially available rapid DNA instruments. A blind test initiated by a DNA expert group showed that the system can correctly produce DNA profiles with 97.29% genotype concordance with standard bench-processing methods, and the profiles can be uploaded into the national DNA database. These results demonstrated that the Quick TargSeq system can rapidly generate reliable DNA profiles in an automated manner and has the potential for use in the field and forensic laboratories.
短串联重复序列（STRs）分析是全球通用的人类身份识别标准方法。插入/缺失多态性（DIPs）可用于生物地理祖先推断。当前的DNA分型需要训练有素的法医工作人员在受控的实验室环境中操作多种专业仪器，这需要6 - 8小时。我们开发了Quick TargSeq 1.0集成系统（以下简称Quick TargSeq），用于从口腔拭子样本和血迹中自动生成STR和DIP图谱。该系统利用微流控生物芯片技术完全集成了DNA提取、聚合酶链反应（PCR）扩增和电泳分离过程。根据DNA分析方法科学工作组的指南，使用RTyper 21或DIP 38芯片盒和单源参考样本进行了内部验证研究。这些结果表明，Quick TargSeq系统能够在大约2小时内处理参考样本并生成STR或DIP图谱，且这些图谱与使用传统STR或DIP分析方法确定的图谱一致。因此，从参考样本中获得了可重复且一致的DNA图谱。在整个研究过程中，未观察到泳道间或批次间的污染。Quick TargSeq系统能从至少擦拭八次的口腔拭子、带有两个2毫米圆盘的干血斑卡或10纳克纯化DNA中生成完整图谱。潜在的PCR抑制剂（即咖啡、吸烟烟草和嚼烟）似乎并未影响该仪器的扩增反应。153个样本的总体成功率和一致性率分别为94.12%和93.44%，与其他市售快速DNA仪器相当。一个DNA专家组发起的盲测表明，该系统能够正确生成DNA图谱，与标准台式处理方法的基因型一致性为97.29%，且这些图谱可上传至国家DNA数据库。这些结果表明，Quick TargSeq系统能够以自动化方式快速生成可靠的DNA图谱，具有在现场和法医实验室使用的潜力。
