论文标题：Silicon µPCR Chip for Forensic STR Profiling with Hybeacon Probe Melting Curves

期刊：

作者：Senne Cornelis, Olivier Tytgat, Maarten Fauvart, Yannick Gansemans, Ann-Sophie Vander Plaetsen, Rodrigo S. Wiederkehr, Dieter Deforce, Filip Van Nieuwerburgh, Tim Stakenborg

发表时间：2019/05/14

数字识别码： 10.1038/s41598-019-43946-5

原文链接：

图1

在中央实验室以外以及犯罪现场进行法明升手机版DNA分析的需求日益增加。如果DNA相关信息的获取时长可以用从几天甚至几周缩短到案发最初的几个小时，会使得一些犯罪调查受益匪浅。然而，由于标准DNA指纹分析法太过复杂且耗时很长，难以实现该分析法的快速化和自动化。

来自比利时根特大学的Dieter Deforce及同事在近期发表于Scientific Reports的一项研究Silicon µPCR Chip for Forensic STR Profiling with Hybeacon Probe Melting Curves中展示了一种利用单片微单晶的技术，该技术有望替代DNA指纹分析法。这项技术将PCR（Polymerase Chain Reaction，聚合酶链反应）扩增技术与硅芯片实验室（LoC, Lab-on-a-chip）中的HyBeacon熔融分析相结合，在快速现场DNA指纹图谱鉴定领域迈出了重要的一步。

图2：芯片正反面图

LoC形体小巧，因而降低了试剂消耗并且增加了便携性。LoC的小型化优势是通过一个集成加热元件实现的，这个集成元件包含24个平行微型反应器，每个反应器的反应体积为0.14L。而高度的平行化使得作者可以同时分析4个短串联重复序列（STR，short tandem repeat）位点以及法明升手机版DNA分析中常见的釉原蛋白性别标记。

如此一来，便可以同时分析对照样本和现场采集的测试样本，并直接进行比较，从而得到更直观的结果。重要的是，通过使用行业标准的半导体制造工艺，可以保证这个仪器的大规模制造。在分析设计和优化之后，可以在芯片上生成完整的5个基因位点信息，测试的结果可以与传统的台式实时PCR仪媲美。在开发商用的、可大规模生产、便携式的法医DNA现场分析型设备进程中，作者的研究是很重要的一步进展。

摘要：The demand to perform forensic DNA profiling outside of centralized laboratories and on the crime scene is increasing. Several criminal investigations would benefit tremendously from having DNA based information available in the first hours rather than days or weeks. However, due to the complexity and time-consuming nature of standard DNA fingerprinting methods, rapid and automated analyses are hard to achieve. We here demonstrate the implementation of an alternative DNA fingerprinting method in a single microchip. By combining PCR amplification and HyBeacon melting assays in a silicon Lab-on-a-chip (LoC), a significant step towards rapid on-site DNA fingerprinting is taken. The small form factor of a LoC reduces reagent consumption and increases portability. Additional miniaturization is achieved through an integrated heating element covering 24 parallel micro-reactors with a reaction volume of 0.14 µl each. The high level of parallelization allows the simultaneous analysis of 4 short tandem repeat (STR) loci and the amelogenin gender marker commonly included in forensic DNA analysis. A reference and crime scene sample can be analyzed simultaneously for direct comparison. Importantly, by using industry-standard semiconductor manufacturing processes, mass manufacturability can be guaranteed. Following assay design and optimization, complete 5-loci profiles could be robustly generated on-chip that are on par with those obtained using conventional benchtop real-time PCR thermal cyclers. Together, our results are an important step towards the development of commercial, mass-produced, portable devices for on-site testing in forensic DNA analysis.

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期刊介绍： () is an online, open access journal from the publishers of Nature. We publish scientifically valid primary research from all areas of the natural and clinical sciences.

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•2-year Median: 2

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