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Rapid sequencing and identification for 18-STRs long amplicon panel using portable devices and nanopore sequencer

文献类型： 外文期刊

作者： Zhang, Jiarong 1 ; Yang, Tingting 1 ; Xie, Zihan 2 ; Ren, Zilin 5 ; Shi, Linyu 1 ; Yan, Jiang-wei 1 ; Ni, Ming 1 ;

作者机构： 1.Shanxi Med Univ, Sch Forens Med, Jinzhong 030600, Peoples R China

2.Acad Mil Med Sci, Beijing 100850, Peoples R China

3.Shanxi Key Lab Forens Med, Jinzhong 030600, Peoples R China

4.Beijing Univ Chem Technol, Sch Life Sci, Beijing 100029, Peoples R China

5.Chinese Acad Agr Sci, Changchun Vet Res Inst, Changchun 130122, Peoples R China

6.Northeast Normal Univ, Sch Informat Sci & Technol, Changchun 130117, Peoples R China

7.Northeast Normal Univ, Inst Computat Biol, Changchun 130117, Peoples R China

关键词： Short tandem repeat (STR); Long-amplicon; Multiplex amplification; Rapid human identification; Nanopore sequencing; Portable devices

期刊名称：GENOMICS （ 影响因子：3.0； 五年影响因子：3.3 ）

ISSN： 0888-7543

年卷期： 2025 年 117 卷 1 期

页码：

收录情况： SCI

摘要： STRs are the most commonly used forensic genetic markers for human identification. Nanopore sequencing has shown the advantages of high portability and large data throughput. Previous studies indicate it has great potential for profiling STRs based on the ligation library preparation method. However, this method, which requires more library preparation time and operations, is unsuitable for rapid STR profiling, particularly for field forensic applications. The transposase-based rapid library preparation method offers the possibility to perform human identification using portable instruments. However, the amplicons of conventional STR panels are too small and would be cut into scraps with rapid methods, making them impractical for genotyping. In this study, we developed an 18-STRs multiplex amplification panel with amplicons of similar to 1.4 Kbp. The PCR conditions were optimized to be finished within 2 h and 12 min, and the PCR products could undergo rapid methods that involved random fragmentation. We found that, on average, 29.16 % of reads from the long-amplicon panel and rapid library kit covered the whole STR region, sufficient for downstream STR profiling analysis. We conducted a small validation experiment on 24 samples using portable instruments powered by a 1.5 kW center dot h portable power source. The entire process took 10.5 h and we obtained enough data from 24 samples to perform trustworthy pairwise identification analysis using the STR profiles. The overall accuracy of the analysis was 95.36 %. In sum, the study evaluated and demonstrated the viability and potential of nanopore sequencing for forensic application in the field.

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