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Rapid DNA analysis for automated processing and interpretation of low DNA content samples

机译：快速的DNA分析可自动处理和解释低DNA含量的样品

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Background Short tandem repeat (STR) analysis of casework samples with low DNA content include those resulting from the transfer of epithelial cells from the skin to an object (e.g., cells on a water bottle, or brim of a cap), blood spatter stains, and small bone and tissue fragments. Low DNA content (LDC) samples are important in a wide range of settings, including disaster response teams to assist in victim identification and family reunification, military operations to identify friend or foe, criminal forensics to identify suspects and exonerate the innocent, and medical examiner and coroner offices to identify missing persons. Processing LDC samples requires experienced laboratory personnel, isolated workstations, and sophisticated equipment, requires transport time, and involves complex procedures. We present a rapid DNA analysis system designed specifically to generate STR profiles from LDC samples in field-forward settings by non-technical operators. By performing STR in the field, close to the site of collection, rapid DNA analysis has the potential to increase throughput and to provide actionable information in real time. Results A Low DNA Content BioChipSet (LDC BCS) was developed and manufactured by injection molding. It was designed to function in the fully integrated Accelerated Nuclear DNA Equipment (ANDE) instrument previously designed for analysis of buccal swab and other high DNA content samples (Investigative Genet. 4(1):1–15, 2013). The LDC BCS performs efficient DNA purification followed by microfluidic ultrafiltration of the purified DNA, maximizing the quantity of DNA available for subsequent amplification and electrophoretic separation and detection of amplified fragments. The system demonstrates accuracy, precision, resolution, signal strength, and peak height ratios appropriate for casework analysis. Conclusions The LDC rapid DNA analysis system is effective for the generation of STR profiles from a wide range of sample types. The technology broadens the range of sample types that can be processed and minimizes the time between sample collection, sample processing and analysis, and generation of actionable intelligence. The fully integrated Expert System is capable of interpreting a wide range or sample types and input DNA quantities, allowing samples to be processed and interpreted without a technical operator.
机译：背景技术对DNA含量低的个案样品进行的短串联重复序列（STR）分析包括那些由于上皮细胞从皮肤转移到物体（例如水瓶上的细胞或帽沿上的细胞），血溅斑，和小的骨头和组织碎片。低DNA含量（LDC）样品在各种环境中都很重要，包括灾难响应小组以帮助受害者识别和家庭团聚，军事行动以识别朋友或敌人，刑事司法鉴定以识别嫌疑人并释放无辜者，以及医学检查员和验尸官办公室以识别失踪人员。处理LDC样品需要经验丰富的实验室人员，孤立的工作站和复杂的设备，需要运输时间，并且涉及复杂的程序。我们提出了一种快速DNA分析系统，该系统专门设计用于由非技术人员在现场转发设置中从LDC样品生成STR谱图。通过在靠近采集地点的现场进行STR，快速的DNA分析有可能提高通量并实时提供可操作的信息。结果通过注射成型开发并生产了低DNA含量生物芯片组（LDC BCS）。它设计用于在完全集成的加速核DNA设备（ANDE）仪器中运行，该仪器以前是为分析口腔拭子和其他高DNA含量的样品而设计的（Investigative Genet。4（1）：1-15，2013）。 LDC BCS执行有效的DNA纯化，然后对纯化的DNA进行微流超滤，从而使可用于后续扩增，电泳分离和检测扩增片段的DNA量最大化。该系统演示了适用于案例分析的准确性，精度，分辨率，信号强度和峰高比。结论LDC快速DNA分析系统可有效地从多种样品类型生成STR谱图。该技术拓宽了可以处理的样品类型的范围，并最大程度地缩短了样品收集，样品处理和分析以及生成可行情报之间的时间。完全集成的专家系统能够解释广泛的或样品类型以及输入的DNA数量，从而无需技术操作员即可处理和解释样品。

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