Developmental Validation of the Microreader 23HS Plex ID System: A Novel Supplementary Non-CODIS STR Multiplex Assay for Forensic Application.

IF 3 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS
Hui Li, He Ren, Fan Yang, Man Chen, Weifen Sun, Lei Jiang, Zhixiao Gao, Yacheng Liu, Xiling Liu
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引用次数: 0

Abstract

A novel supplementary non-CODIS STR multiplex assay designated as the "Microreader 23HS Plex ID System" was developed. The Microreader 23HS Plex ID System enables simultaneous profiling of 23 STR loci and the amelogenin locus. The majority of these loci are non-CODIS STRs (D4S2408, D9S2157, D20S161, D3S2459, D18S1364, D13S305, D1S2142, D19S400, D6S1017, D7S1517, D2S1776, D2S1360, D3S1744, D16S3391, D3S1545, D11S4463, D20S85, D1S549, D10S2325, D21S2055), with the exception of three CODIS STRs (D2S441, D12S391, and D22S1045). Followed the recommendations of Scientific Working Group on DNA Analysis Methods (SWGDAM) and the Chinese validation standards, a comprehensive set of validation studies were conducted, encompassing PCR conditions, stutter ratio and peak height balance, sensitivity, precision and accuracy, reproducibility, species specificity, inhibition, as well as mixture testing. The results demonstrated that the Microreader 23HS Plex ID System is a reliable and robust assay, with well-balanced peak heights, high precision and accuracy, species specificity, and resistance to common inhibitors. The sensitivity of the assay was determined to be 0.125 ng of template DNA. In mixture study, all minor alleles were detected in two-sample mixtures across various ratios (1:19, 1:9, 1:4, 3:7, 2:3, 1:1, 3:2, 4:1, 9:1, and 19:1). In population study, a total of 500 unrelated individuals of Han ethnicity from East China were genotyped. The allele frequencies and forensic population genetic parameters were calculated, with a cumulative random match probability of 7.757 × 10-27, and a total power of discrimination exceeding 0.999,999,999,999,999,999,999,999,99. In conclusion, the Microreader 23HS Plex ID System shows promise as a valuable supplementary tool for forensic applications, particularly in addressing complex kinship testing and challenges posed by STR mutation.

Microreader 23HS Plex ID 系统的开发验证:用于法医应用的新型补充性非 CODIS STR 多重检测。
我们开发了一种新型的辅助性非 CODIS STR 多重检测方法,命名为 "Microreader 23HS Plex ID 系统"。Microreader 23HS Plex ID 系统可同时分析 23 个 STR 基因座和淀粉样蛋白基因座。这些基因位点大部分是非 CODIS STR(D4S2408、D9S2157、D20S161、D3S2459、D18S1364、D13S305、D1S2142、D19S400、D6S1017、D7S1517、D2S1776、D2S1360、D3S1744、D16S1776、D3S1744)、D3S1744, D16S3391, D3S1545, D11S4463, D20S85, D1S549, D10S2325, D21S2055),只有三个 CODIS STR(D2S441、D12S391 和 D22S1045)除外。根据 DNA 分析方法科学工作组(SWGDAM)的建议和中国的验证标准,我们进行了一套全面的验证研究,包括 PCR 条件、滞后比和峰高平衡、灵敏度、精密度和准确度、重现性、物种特异性、抑制以及混合测试。结果表明,Microreader 23HS Plex ID 系统是一种可靠、稳健的检测方法,峰高均衡、精密度和准确度高、物种特异性强,而且对常见抑制剂具有抗性。测定灵敏度为 0.125 纳克模板 DNA。在混合物研究中,在不同比例(1:19、1:9、1:4、3:7、2:3、1:1、3:2、4:1、9:1 和 19:1)的两个样本混合物中检测到了所有小等位基因。在人群研究中,共对华东地区 500 名无血缘关系的汉族个体进行了基因分型。计算了等位基因频率和法医人群遗传参数,累计随机匹配概率为 7.757 × 10-27,总鉴别力超过 0.9999999999999999999。总之,Microreader 23HS Plex ID 系统有望成为法医应用的重要辅助工具,特别是在解决复杂的亲属关系测试和 STR 变异带来的挑战方面。
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来源期刊
ELECTROPHORESIS
ELECTROPHORESIS 生物-分析化学
CiteScore
6.30
自引率
13.80%
发文量
244
审稿时长
1.9 months
期刊介绍: ELECTROPHORESIS is an international journal that publishes original manuscripts on all aspects of electrophoresis, and liquid phase separations (e.g., HPLC, micro- and nano-LC, UHPLC, micro- and nano-fluidics, liquid-phase micro-extractions, etc.). Topics include new or improved analytical and preparative methods, sample preparation, development of theory, and innovative applications of electrophoretic and liquid phase separations methods in the study of nucleic acids, proteins, carbohydrates natural products, pharmaceuticals, food analysis, environmental species and other compounds of importance to the life sciences. Papers in the areas of microfluidics and proteomics, which are not limited to electrophoresis-based methods, will also be accepted for publication. Contributions focused on hyphenated and omics techniques are also of interest. Proteomics is within the scope, if related to its fundamentals and new technical approaches. Proteomics applications are only considered in particular cases. Papers describing the application of standard electrophoretic methods will not be considered. Papers on nanoanalysis intended for publication in ELECTROPHORESIS should focus on one or more of the following topics: • Nanoscale electrokinetics and phenomena related to electric double layer and/or confinement in nano-sized geometry • Single cell and subcellular analysis • Nanosensors and ultrasensitive detection aspects (e.g., involving quantum dots, "nanoelectrodes" or nanospray MS) • Nanoscale/nanopore DNA sequencing (next generation sequencing) • Micro- and nanoscale sample preparation • Nanoparticles and cells analyses by dielectrophoresis • Separation-based analysis using nanoparticles, nanotubes and nanowires.
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