Skip to main content
SpringerLink
Log in

Development and validation of novel 8-dye short tandem repeat multiplex system for forensic applications

  • Original Article
  • Published:
International Journal of Legal Medicine Aims and scope Submit manuscript

Abstract

DNA profiling of short tandem repeats (STRs) is the primary method for genotyping forensic samples. However, degraded DNA and trace samples are still major problems for commercial 5- or 6-dye STR kits. In order to improve the performance of this method, we developed a novel 8-dye STR multiplex system containing 18 autosomal loci (D3S1358, D1S1656, TPOX, D16S539, vWA, D6S1043, D2S1338, CSF1PO, D19S433, D7S820, FGA, D8S1179, D5S818, D13S317, TH01, D21S11, D12S391, and PentaD) and the sex-determining locus Amelogenin, with all fragments smaller than 330 bases. Validation was carried out as recommended by the Scientific Working Group on DNA Analysis Methods. The results showed that complete profiles were obtainable when the input DNA was as low as 0.0625 ng. Full profiles were obtained even in the presence of inhibitors such as humic acid (< 300 ng/μl), hematin (< 100 μM), and indigo (0.01%). The 8-dye STR multiplex system also showed good performance in the detection degraded DNA samples. These results indicate that the 8-dye STR multiplex system is suitable for human DNA genotyping, including for difficult forensic materials.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Butler JM (2005) Forensic DNA typing: biology, technology, and genetics of STR markers, 2nd edn. Elsevier, New York

    Google Scholar 

  2. Holmes AS, Roman MG, Hughes-Stamm S (2018) In-field collection and preservation of decomposing human tissues to facilitate rapid purification and STR typing. Forensic Sci Int Genet 36:124–129. https://doi.org/10.1016/j.fsigen.2018.06.015

    Article  CAS  PubMed  Google Scholar 

  3. Oostdik K, Lenz K, Nye J, Schelling K, Yet D, Bruski S, Strong J, Buchanan C, Sutton J, Linner J, Frazier N, Young H, Matthies L, Sage A, Hahn J, Wells R, Williams N, Price M, Koehler J, Staples M, Swango KL, Hill C, Oyerly K, Duke W, Katzilierakis L, Ensenberger MG, Bourdeau JM, Sprecher CJ, Krenke B, Storts DR (2014) Developmental validation of the PowerPlex (®) Fusion System for analysis of casework and reference samples: a 24-locus multiplex for new database standards. Forensic Sci Int Genet 12:69–76. https://doi.org/10.1016/j.fsigen.2014.04.013

    Article  CAS  PubMed  Google Scholar 

  4. Ensenberger MG, Lenz KA, Matthies LK, Hadinoto GM, Schienman JE, Przech AJ, Morganti MW, Renstrom DT, Baker VM, Gawrys KM, Hoogendoorn M, Steffen CR, Martín P, Alonso A, Olson HR, Sprecher CJ, Storts DR (2016) Developmental validation of the PowerPlex (®) Fusion 6C System. Forensic Sci Int Genet 21:134–144. https://doi.org/10.1016/j.fsigen.2015.12.011

    Article  CAS  PubMed  Google Scholar 

  5. Mulero JJ, Chang CW, Lagacé RE, Wang DY, Bas JL, McMahon TP, Hennessy LK (2008) Development and validation of the AmpFlSTR MiniFiler PCR amplification kit: a MiniSTR multiplex for the analysis of degraded and/or PCR inhibited DNA. J Forensic Sci 53(4):838–852. https://doi.org/10.1111/j.1556-4029.2008.00760.x

    Article  CAS  PubMed  Google Scholar 

  6. Drábek J, Chung DT, Butler JM, McCord BR (2004) Concordance study between miniplex assays and a commercial STR typing kit. J Forensic Sci 49(4):859–860

    Article  Google Scholar 

  7. Liu QL, Luo H, Zhao H, Huang XL, Cheng JD, Lu DJ (2014) Recombination analysis of autosomal short tandem repeats in Chinese Han families. Electrophoresis 35(6):883–887. https://doi.org/10.1002/elps.201300417

    Article  CAS  PubMed  Google Scholar 

  8. Wu W, Hao H, Liu Q, Han X, Wu Y, Cheng J, Lu D (2014) Analysis of linkage and linkage disequilibrium for syntenic STRs on 12 chromosomes. Int J Legal Med 128(5):735–739. https://doi.org/10.1007/s00414-014-1032-y

    Article  PubMed  Google Scholar 

  9. Scientific Working Group on DNA Analysis Methods (SWGDAM). Validation guidelines for DNA analysis methods, https://www.swgdam.org

  10. Wang L, Zhao XC, Ye J, Liu JJ, Chen T, Bai X, Zhang J, Ou Y, Hu L, Jiang BW, Wang F (2014) Construction of a library of cloned short tandem repeat (STR) alleles as universal templates for allelic ladder preparation. Forensic Sci Int Genet 12:136–143. https://doi.org/10.1016/j.fsigen.2014.06.005

    Article  CAS  PubMed  Google Scholar 

  11. Sharma V, van der Plaat DA, Liu Y, Wurmbach E (2020) Analyzing degraded DNA and challenging samples using the ForenSeqTM DNA Signature Prep kit. Science & justice: journal of the Forensic Science Society 60(3):243–252. https://doi.org/10.1016/j.scijus.2019.11.004

    Article  Google Scholar 

  12. Jiang B, He W, Jin C, Liu Y, Wen D, Wang C, Zeye MMJ, Li J, Zha L (2021) Developmental validation of the STRscan-17LC kit: a 6 Dye STR kit enhanced stability and ability to detect degraded samples. Int J Legal Med 135(2):431–440. https://doi.org/10.1007/s00414-020-02490-y

    Article  PubMed  Google Scholar 

  13. Zhao F, Wu X, Cai G, Xu, C (2003) The application of Mdified-Powerstates software in forensic biostatistics (In Chinese). Chinese Journal of Forensic Medicine. 18:297–298

  14. Wang DY, Chang CW, Lagacé RE, Calandro LM, Hennessy LK (2012) Developmental validation of the AmpFℓSTR® Identifiler® Plus PCR amplification kit: an established multiplex assay with improved performance. J Forensic Sci 57(2):453–465. https://doi.org/10.1111/j.1556-4029.2011.01963.x

    Article  CAS  PubMed  Google Scholar 

  15. Monroe C, Grier C, Kemp BM (2013) Evaluating the efficacy of various thermo-stable polymerases against co-extracted PCR inhibitors in ancient DNA samples. Forensic Sci Int 228(1–3):142–153. https://doi.org/10.1016/j.forsciint.2013.02.029

    Article  CAS  PubMed  Google Scholar 

  16. Opel KL, Chung D, McCord BR (2010) A study of PCR inhibition mechanisms using real time PCR. J Forensic Sci 55(1):25–33. https://doi.org/10.1111/j.1556-4029.2009.01245.x

    Article  CAS  PubMed  Google Scholar 

  17. Li J, Luo H, Song F, Zhang L, Deng C, Yu Z, Gao T, Liao M, Hou Y (2017) Validation of the MicroreaderTM 23sp ID system: a new STR 23-plex system for forensic application. Forensic Sci Int Genet 27:67–73. https://doi.org/10.1016/j.fsigen.2016.12.005

    Article  CAS  PubMed  Google Scholar 

  18. Kraemer M, Prochnow A, Bussmann M, Scherer M, Peist R, Steffen C (2017) Developmental validation of QIAGEN Investigator® 24plex QS Kit and Investigator® 24plex GO! Kit: two 6-dye multiplex assays for the extended CODIS core loci. Forensic Sci Int Genet 29:9–20. https://doi.org/10.1016/j.fsigen.2017.03.012

    Article  CAS  PubMed  Google Scholar 

  19. Bai X, Yao Y, Wang C, Li W, Wang Y, Song W, Mo X, Ma W, Ye J (2019) Development of a new 25plex STRs typing system for forensic application. Electrophoresis 40(12–13):1662–1676. https://doi.org/10.1002/elps.201900021

    Article  CAS  PubMed  Google Scholar 

  20. Zhang J, Zhang J, Tao R, Jiang L, Chen L, Li X, Li C, Zhang S (2020) A newly devised multiplex assay of novel polymorphic non-CODIS STRs as a valuable tool for forensic application. Forensic Sci Int Genet 48:102341. https://doi.org/10.1016/j.fsigen.2020.102341

    Article  CAS  PubMed  Google Scholar 

  21. Wang S, Song F, Xie M, Zhang K, Xie B, Huang Z, Luo H (2020) Evaluation of a six-dye multiplex composed of 27 markers for forensic analysis and databasing. Mol Genet Genomic Med 8(9):e1419. https://doi.org/10.1002/mgg3.1419

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Li S, Chen L, Wang Y, Xu Q, Liu H, Li Y, Liu C (2018) Developmental validation of a 6-dye STR kit with 27 loci. Int J Legal Med 132(2):335–342. https://doi.org/10.1007/s00414-017-1586-6

    Article  PubMed  Google Scholar 

  23. Yang M, Yin C, Lv Y, Yang Y, Chen J, Yu Z, Liu X, Xu M, Chen F, Wu H, Yan J (2016) Development of a rapid 21-plex autosomal STR typing system for forensic applications. Electrophoresis 37(21):2789–2799. https://doi.org/10.1002/elps.201500498

    Article  CAS  PubMed  Google Scholar 

  24. Wang Z, Zhou D, Jia Z, Li L, Wu W, Li C, Hou Y (2016) Developmental validation of the Huaxia Platinum System and application in 3 main ethnic groups of China. Sci Rep 6:31075. https://doi.org/10.1038/srep31075

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Han J, Sun J, Zhao L, Zhao W, Liu Y, Li C (2017) Validation study of a 15-plex rapid STR amplification system for human identification. Forensic Sci Int Genet 28:71–81. https://doi.org/10.1016/j.fsigen.2017.01.015

    Article  CAS  PubMed  Google Scholar 

  26. Ludeman MJ, Zhong C, Mulero JJ, Lagacé RE, Hennessy LK, Short ML, Wang DY (2018) Developmental validation of GlobalFilerTM PCR amplification kit: a 6-dye multiplex assay designed for amplification of casework samples. Int J Legal Med 132(6):1555–1573. https://doi.org/10.1007/s00414-018-1817-5

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Funding

This project was supported by the National Natural Science Foundation of China (NSFC, No. 81871533) and the Natural Science Foundation of Hunan Province (No. 2020JJ4779).

This research was also supported by the foundation from joint postgraduate training base between Central South University and the First Research Institute of the Ministry of Public Security of P.R.C (No. 2020pyjd23).

Author information

Authors and Affiliations

  1. The First Research Institute of the Ministry of Public Security of P.R.C, Haidian District, No.1. Shouti South Road, Beijing, 100044, China

    Bowei Jiang, Feng Wang, Liming Zhang, Haibo Rong, Tao Zhang, Chuan Jin & Li Chen

  2. Department of Forensic Science, School of Basic Medical Sciences, Central South University, Hunan Province, No. 172. Tongzipo Road, Changsha, 410013, China

    Weifeng Qu, Jienan Li, Dan Wen, Moutanou Modeste Judes Zeye, Wei He, Chudong Wang, Hao Xing, Ying Liu, Jifeng Cai & Lagabaiyila Zha

Authors
  1. Bowei Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Weifeng Qu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Feng Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Liming Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Haibo Rong
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jienan Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Dan Wen
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Moutanou Modeste Judes Zeye
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Wei He
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Chudong Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Hao Xing
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Tao Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Chuan Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Li Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Ying Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Jifeng Cai
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Lagabaiyila Zha
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lagabaiyila Zha.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Additional information

Publisher's note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Bowei Jiang and Weifeng Qu contributed equally to this article and shared first authorship

Bowei Jiang and Weifeng Qu contributed equally to this article and shared first authorship.

Supplementary Information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jiang, B., Qu, W., Wang, F. et al. Development and validation of novel 8-dye short tandem repeat multiplex system for forensic applications. Int J Legal Med 135, 2263–2274 (2021). https://doi.org/10.1007/s00414-021-02695-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00414-021-02695-9

Keywords

Search

Navigation