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Evaluation of the MHSeqTyper47 kit for forensically challenging DNA samples

  • Author Footnotes
    1 These authors contributed equally to this work.
    Yao-Sen Feng
    Footnotes
    1 These authors contributed equally to this work.
    Affiliations
    National Engineering Laboratory for Forensic Science, Key Laboratory of Forensic Genetics of Ministry of Public Security, Institute of Forensic Science, Ministry of Public Security, Beijing 100038, China
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  • Author Footnotes
    1 These authors contributed equally to this work.
    Chi Zhang
    Footnotes
    1 These authors contributed equally to this work.
    Affiliations
    National Engineering Laboratory for Forensic Science, Key Laboratory of Forensic Genetics of Ministry of Public Security, Institute of Forensic Science, Ministry of Public Security, Beijing 100038, China
    Search for articles by this author
  • Qing-Feng Chen
    Affiliations
    National Engineering Laboratory for Forensic Science, Key Laboratory of Forensic Genetics of Ministry of Public Security, Institute of Forensic Science, Ministry of Public Security, Beijing 100038, China
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  • Yu Wang
    Affiliations
    National Engineering Laboratory for Forensic Science, Key Laboratory of Forensic Genetics of Ministry of Public Security, Institute of Forensic Science, Ministry of Public Security, Beijing 100038, China
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  • Ke-Lai Kang
    Affiliations
    National Engineering Laboratory for Forensic Science, Key Laboratory of Forensic Genetics of Ministry of Public Security, Institute of Forensic Science, Ministry of Public Security, Beijing 100038, China
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  • Jie Zhao
    Affiliations
    National Engineering Laboratory for Forensic Science, Key Laboratory of Forensic Genetics of Ministry of Public Security, Institute of Forensic Science, Ministry of Public Security, Beijing 100038, China
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  • An-Quan Ji
    Correspondence
    Corresponding authors.
    Affiliations
    National Engineering Laboratory for Forensic Science, Key Laboratory of Forensic Genetics of Ministry of Public Security, Institute of Forensic Science, Ministry of Public Security, Beijing 100038, China
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  • Jian Ye
    Correspondence
    Corresponding authors.
    Affiliations
    National Engineering Laboratory for Forensic Science, Key Laboratory of Forensic Genetics of Ministry of Public Security, Institute of Forensic Science, Ministry of Public Security, Beijing 100038, China
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  • Le Wang
    Correspondence
    Corresponding authors.
    Affiliations
    National Engineering Laboratory for Forensic Science, Key Laboratory of Forensic Genetics of Ministry of Public Security, Institute of Forensic Science, Ministry of Public Security, Beijing 100038, China
    Search for articles by this author
  • Author Footnotes
    1 These authors contributed equally to this work.

      Highlights

      • The MHSeqTyper47 kit, which covered 47 microhaplotypes, was evaluated for forensic use.
      • MiSeq FGx sequencing metrics for seven runs of MHSeqTyper47 sequencing were reported.
      • The MHSeqTyper47 data was accurate, repeatable, inter-locus and intra-locus balanced.
      • Sensitivity, degraded DNA, PCR inhibition, and case-type studies were performed.
      • Microhaplotypes exhibited an advantage over CE-STR in mixture interpretation.

      Abstract

      Microhaplotypes have been highly regarded for forensic mixture DNA deconvolution because they do not experience interference from stutters in the same way as short tandem repeat markers, and they tend to be more polymorphic than single nucleotide polymorphism markers. However, forensic microhaplotype kits have not been reported. The MHSeqTyper47 kit genotypes 47 microhaplotype loci. In this study, MiSeq FGx sequencing metrics for MHSeqTyper47 were presented, and the genotyping accuracy of this kit was examined. The sensitivity of MHSeqTyper47 reached 62.5 pg, and full genotyping results were obtained from degraded DNA samples with degradation indexes ≤ 3.00. Full genotypes were obtained in the presence of 100 ng/μL tannin, 50 μM heme, 25 ng/μL humic acid, and 1.25 μg/μL indigo dye. In DNA mixture studies, a minimum of 31 loci of the minor contributor were correctly genotyped at 1:99 or 99:1 mixing ratios, with the cumulative random matching probability of these loci reaching 4.54 × 10−25. Mixing ratios could be reliably predicted from two-donor DNA mixtures based on the loci with four called alleles. Taken together, these data showed that the MHSeqTyper47 kit was effective for forensically challenging DNA analysis.

      Keywords

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