Internal validation study of a newly developed 24-plex Y-STRs genotyping system for forensic application

Abstract

Prior to implementing a new kit into application, developmental validation should be conducted to demonstrate the robustness and applicability of the kit. In this study, 24 Y-STR loci from the AGCU Y SUPP STR kit were tested including 11 loci overlapping with other commercial kits (DYS385a/b, DYS635, DYS533, DYS481, DYS549, DYS460, DYS527a/b, DYS522, and DYS444) and 13 new loci (DYS531, DYS630, DYS622, DYS552, DYS510, DYS459a/b, DYS446, DYS443, DYS587, Y-GATA-A10, DYS520, and DYS557). Developmental validation including PCR-related studies, sensitivity, stability, and species specificity studies were conducted. The performance of the kit in genotyping case-type samples was also estimated. The results indicated that the kit is robust, accurate and sensitive and is able to detect male samples without being affected by female samples or other species. Population data were obtained with this kit in Chinese Xibe group as well. Totally 139 different haplotypes were obtained from 167 male samples and demonstrated that this typing system is relatively discriminative.

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Funding

This project was supported by the Fundamental Research Funds for the Central Universities, China (No. xjj2018165), the Key Project for Science Research and Development of Shaanxi Province (2018SF-119), the National Natural Science Foundation of China (No. 81525015) and the Guangdong Province Universities and Colleges Pearl River Scholar Funded Scheme (GDUPS, 2017) .

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Correspondence to Bofeng Zhu.

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Electronic supplementary material

ESM 1

. Genotyping results of PCR reaction component study in which the concentration of Reaction Mix varied from 2.0–6.0 μL. (PDF 124 kb)

ESM 2

. Genotyping results of PCR reaction component study in which the concentration of Y SUPP Primers varied from 1.0–3.0 μL. (PDF 100 kb)

ESM 3

. Genotyping results of PCR reaction component study in which the concentration of C-Taq polymerase varied from 0.2–0.6 μL. (PDF 142 kb)

ESM 4

. Genotyping results of thermal cycling parameter study in which the cycle number varied from 28-32. (PDF 143 kb)

ESM 5

. Genotyping results of thermal cycling parameter study in which the annealing temperature varied from 57-64 °C. (PDF 145 kb)

ESM 6

. Genotyping results of Y-GATA-A10, DYS520 and DYS443 loci when final extension time varied from 0-60 min. (PDF 245 kb)

ESM 7

. Profile of Male Control DNA 9948 created using the optimal conditions (the same as the manufacture’s instruction) (PDF 446 kb)

ESM 8

. Genotyping results when total reaction volumes varied from 6.25–25 μL. (PDF 124 kb)

ESM 9

. Genotyping results of species specificity studies. (PDF 173 kb)

ESM 10

. Genotyping results of hair, blood stain, oral swab and saliva card samples from three males. (PDF 288 kb)

ESM 11

. Allelic frequencies and GD values of the 24 Y-STR loci in 167 Xibe male individuals from Xinjiang (XLSX 13 kb)

ESM 12

. The 139 different haplotypes obtained at the 24 Y-STR loci in 167 Xibe male individuals from Xinjiang (XLSX 24 kb)

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Meng, H., Guo, Y., Jin, X. et al. Internal validation study of a newly developed 24-plex Y-STRs genotyping system for forensic application. Int J Legal Med 133, 733–743 (2019). https://doi.org/10.1007/s00414-019-02028-x

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Keywords

  • Validation
  • Y-STR
  • Forensic science
  • Genotyping
  • Xibe ethnic group