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Developmental validation of the Microreader™ RM-Y ID System: a new rapidly mutating Y-STR 17-plex system for forensic application

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Abstract

Y-chromosomal short tandem repeats (Y-STRs) are widely applied to evolutionary, genealogical, and kinship analyses of male linages in forensic studies, but these low to midrange mutated Y-STRs typically fail to separate related males from the same paternal lineage. Recently, rapidly mutating Y-STRs (RM Y-STRs) have been demonstrated to improve the differentiation of male relatives and individuals. The Microreader™ RM-Y ID System is a new RM Y-STR kit that is capable of simultaneously amplifying 17 RM Y-STRs. Herein, to verify the efficiency and accuracy of the Microreader™ RM-Y ID System, developmental validation was conducted, including PCR-based studies, sensitivity, stability, species specificity, mixture, stutter percentage, and precision studies. Full profiles could be obtained when the hematin concentration was 250 μM, humic acid concentration was 1500 ng/μl, and tannic acid concentration was 200 ng/μl. Full profiles of the mixture of males/males could be detected up to a ratio of 19:1, and full profiles of females/males could always be detected even at ratios up to 24,000:1. Moreover, the forensic characteristics of 250 DNA-confirmed father-son pairs were analysed. The results showed that these 17 RM Y-STRs had high power for forensic discrimination (HD = 1) in the Chinese Han population, and the mutation rates were in the range of 4 × 10−3 (95% CI 1.00 × 10−4 to 2.21 × 10−2, DYS464) to 8.8 × 10−2 (95% CI 5.60 × 10−2 to 1.30 × 10−1, DYF399S1), indicating that the kit was effective for RM Y-STR studies and absolute individualisation of interrelated male individuals.

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Funding

This work was funded by the National Natural Science Foundation of China, grant number 81772030.

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Authors and Affiliations

  1. Department of Forensic Genetics, West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, 3-16 Renmin South Road, Chengdu, 610041, China

    Yuxiang Zhou, Feng Song, Shuangshuang Wang, Ke Zhang, Xiaowen Wei, Xindi Wang & Haibo Luo

  2. Department of Forensic Pathology, West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu, 610041, China

    Hao Dai

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  1. Yuxiang Zhou
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Correspondence to Haibo Luo.

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Approval was obtained from the ethics committee of Sichuan University. The procedures used in this study adhere to the tenets of the Declaration of Helsinki.

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Yuxiang Zhou and Feng Song contributed equally to this work and should be considered co-first authors.

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Zhou, Y., Song, F., Dai, H. et al. Developmental validation of the Microreader™ RM-Y ID System: a new rapidly mutating Y-STR 17-plex system for forensic application. Int J Legal Med 136, 501–512 (2022). https://doi.org/10.1007/s00414-021-02632-w

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