Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Sep;8(9):e1419.
doi: 10.1002/mgg3.1419. Epub 2020 Jul 16.

Evaluation of a six-dye multiplex composed of 27 markers for forensic analysis and databasing

Shuangshuang Wang  1 , Feng Song  1 , Mingkun Xie  2 , Ke Zhang  1 , Bowen Xie  1 , Zhanglong Huang  3 , Haibo Luo  1   4
Affiliations

Evaluation of a six-dye multiplex composed of 27 markers for forensic analysis and databasing

Shuangshuang Wang et al. Mol Genet Genomic Med. 2020 Sep.

Retraction in

Abstract

Background: Short tandem repeat (STR) markers play a significant role in genetic applications and have proved to be effective for the personal identification in forensic medicine. In this study, a six-dye multiplex composed of 23 autosomal STR loci (TH01, D3S1358, Penta D, D6S1043, D21S11, TPOX, D1S1656, D12S391, Penta E, D10S1248, D22S1045, D19S433, D8S1179, D2S1338, D2S441, D18S51, vWA, FGA, D16S539, CSF1PO, D13S317, D5S818, D7S820), one Y chromosome STR (DYS391), two internal quality control markers (Quality Sensor QS1 and QS2), and Amelogenin was evaluated.

Methods: Evaluation studies, including PCR-based studies, sensitivity studies, species specificity studies, stability studies, DNA mixture studies, concordance studies, and precision evaluations were performed according to the guidelines of "Validation Guidelines for Forensic DNA Analysis Methods (2016)" by the Scientific Working Group on DNA Analysis Methods (SWGDAM). In addition, the forensic characteristics of 357 unrelated male samples from Han and Hui populations in China were investigated using 27 markers.

Results: Full STR profiles were obtained from different reaction volumes (5 ~ 25 μl), cycle numbers (28 ~ 34 cycles) and annealing temperatures (58 ~ 62°C). All STR profiles were obtained at humic acid concentration of up to 200 ng/μl and hematin concentration of up to 500 μM. No peaks were observed in most common animal samples except two innovative internal PCR controls (Quality Sensor QS1 and QS2). The six-dye multiplex showed a notably high value for the combined probability of exclusion (CPE), exhibiting values of with 0.99999999977688 in the Han population and 0.999999999583875 in the Hui population. The values of combined probability of discrimination (CPD) were 0.999999999999999999999999999997453 in the Han population and 0. 999999999999999999999999999994398 in the Hui population. In addition, concordance studies showed that there was no difference with the AGCU Express Marker 22 Kit.

Conclusion: The results indicated that the Investigator® 26plex QS Kit is a robust, reliable, and suitable tool for forensic analysis and databasing.

Keywords: concordance study; forensic science; mixture study; sensitivity; short tandem repeat (STR).

PubMed Disclaimer

Conflict of interest statement

No potential conflict of interest was reported by the authors.

Figures

Figure 1
Figure 1
Arrangement of loci of the Investigator® 26plex QS Kit
Figure 2
Figure 2
Results of the sensitivity studies. The concentration of template DNA ranged from 1 ng to 0.03125 ng
Figure 3
Figure 3
Results of the mixture studies. A total of 1 ng template DNA mixed with 9947A and 9948 at various ratios (1:1, 2: 1,4:1, 9:1, 19:1) was tested
Figure 4
Figure 4
Sizing variation of all allelic ladders performed on a 3500 Genetic Analyzer (n = 22)
See this image and copyright information in PMC

References

    1. Bender, K. , Farfan, M. J. , & Schneider, P. M. (2004). Preparation of degraded human DNA under controlled conditions. Forensic Science International, 139(2–3), 135–140. 10.1016/j.forsciint.2003.10.003 - DOI - PubMed
    1. Geng, T. , & Mathies, R. A. (2015). Minimizing inhibition of PCR‐STR typing using digital agarose droplet microfluidics. Forensic Science International: Genetics, 14, 203–209. 10.1016/j.fsigen.2014.10.007 - DOI - PubMed
    1. Gibson‐Daw, G. , Crenshaw, K. , & McCord, B. (2018). Optimization of ultrahigh‐speed multiplex PCR for forensic analysis. Analytical and Bioanalytical Chemistry, 410(1), 235–245. 10.1007/s00216-017-0715-x - DOI - PubMed
    1. Jobling, M. A. , & Gill, P. (2004). Encoded evidence: DNA in forensic analysis. Nature Reviews Genetics, 5(10), 739–751. 10.1038/nrg1455 - DOI - PubMed
    1. Kowalczyk, M. , Zawadzka, E. , Szewczuk, D. , Gryzinska, M. , & Jakubczak, A. (2018). Molecular markers used in forensic genetics. Medicine, Science and the Law, 58(4), 201–209. 10.1177/0025802418803852 - DOI - PubMed

Publication types