International Journal of Legal Medicine

, Volume 132, Issue 2, pp 335–342 | Cite as

Developmental validation of a 6-dye STR kit with 27 loci

  • Shuanglin Li
  • Ling Chen
  • Yanfang Wang
  • Quyi Xu
  • Hong Liu
  • Yue Li
  • Chao Liu
Original Article

Abstract

The EX27 System is a 6-dye (i.e., FAM, HEX, TAMAR, ROX, VIG, and SIZ-500) kit that combines the 24 autosomal STR loci (i.e., D3S1358, D13S317, D7S820, D16S539, D1S1656, Penta E, TPOX, TH01, D2S1338, CSF1PO, Penta D, D10S1248, D19S433, vWA, D21S11, D18S51, D6S1043, D8S1179, D5S818, D12S391, FGA, D22S1045, D2S441, SE33) with a Y-STR locus (DYS391), an Indel site (YIndel) and Amelogenin, which is designed to amplify DNA from extracts as well as direct amplification from substrates. The new system contains most of the widely used autosomal STR loci around the world and is sensitive to the male component in mixtures and is more accurate in gender determination owing to the DYS391 and YIndel.

To demonstrate the suitability for personal identification and parentage testing, the system was validated for sensitivity of detection, species specificity, inhibitor tolerance, DNA mixtures, reproducibility study, and concordance studies based on the SWGDAM developmental validation guidelines. In view of the results, the EX27 System is a robust and reliable amplification kit which can be used for human identification testing.

Keywords

Forensic Science Short tandem repeat (STR) 6-dye multiplexing system 

Notes

Acknowledgements

The authors would like to thank the donors who generously provide samples for this validation study and thank support from the Ministry of Public Security application innovation projects (NO. 2011YYCXGDST076) as well as Science and Technology project of Guangdong Province (NO. 2010A060801001).

Supplementary material

414_2017_1586_MOESM1_ESM.docx (1.8 mb)
ESM 1 (DOCX 1877 kb)
414_2017_1586_MOESM2_ESM.docx (15 kb)
ESM 2 (DOCX 15 kb)
414_2017_1586_MOESM3_ESM.docx (611 kb)
ESM 3 (DOCX 610 kb)

References

  1. 1.
    Hares DR (2012) Expanding the CODIS core loci in the United States. Forensic Sci Int Genet 6(1):e52–e54CrossRefPubMedGoogle Scholar
  2. 2.
    Tozzo P, Giuliodori A, Corato S, Ponzano E, Rodriguez D, Caenazzo L (2013) Deletion of amelogenin Y-locus in forensics: literature revision and description of a novel method for sex cnfirmation. J Forensic Legal Med 20(5):387–391CrossRefGoogle Scholar
  3. 3.
    Scientific Working Group on DNA Analysis Methods (SWGDAM), Validation Guidelines for DNA Analysis Methods, 2013. Available from: http://swgdam. ord/SWGDAM Validation Guidelines Approved Dec 2012.pdfGoogle Scholar
  4. 4.
    Zhao J, Wang Y, Wang G, Ma Y, Shi H, Wen J, et.al (2015), Application of the microwave digestion-vacuum filtration-automated scanning electron microscopy method fordiatom detection in the diagnosis of drowning. J Forensic Legal Med 33:125–128.Google Scholar
  5. 5.
    Huston K.A.(1998), Statistical analysis of STR data Available from: https://www.promega.com/resources/profiles-in-dna/1998/statistical-analysis-of-str-data
  6. 6.
    Magnuson VL, Ally DS, Nylund SJ, Karanjawala ZE, Rayman JB, Knapp JI, et.al. (1996). Substrate nucleotide-determined non-templated addition of adenine by Taq DNA polymerase: implications for PCR-based genotyping and cloning. BioTechniques 21(4):700–709.Google Scholar
  7. 7.
    Akane A, Matsubara K, Nakamura H, Takahashi S, Kimura K (1994) Identification of the heme compound copurified with deoxyribonucleic acid (DNA) from bloodstains, a major inhibitor of polymerase chain reaction (PCR) amplification. J Forensic Sci 39(2):362–372CrossRefPubMedGoogle Scholar
  8. 8.
    Tsai YL, Olson BH (1992) Detection of low numbers of bacterial cells in soils and sediments by polymerase chain reaction. Appl Environ Microbiol 58(2):754–757PubMedPubMedCentralGoogle Scholar
  9. 9.
    Rossen L, Nørskov P, Holmstrøm K, Rasmussen OF (1992) Inhibition of PCR by components of food samples, microbial diagnostic assays and DNA-extraction solutions. Int J Food Microbiol 17(1):37–45CrossRefPubMedGoogle Scholar
  10. 10.
    Hennessy LK, Mehendale N, Chear K, Jovanovich S, Williams S, Park C, et.al. (2014). Developmental validation of the GlobalFiler ® express kit, a 24-marker STR assay, on the RapidHIT ® system, Forensic Sci Int Genet, 13: 247–258.Google Scholar
  11. 11.
    Ensenberger MG, Lenz KA, Matthies LK, Hadinoto GM, Schienman JE, Przech AJ, et.al. (2016). Developmental validation of the PowerPlex® fusion 6C system. Forensic Sci Int Genet. 21:134–144.Google Scholar
  12. 12.
    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-dyemultiplex assays for the extended CODIS core loci. Forensic Sci Int Genet 29:9–20CrossRefPubMedGoogle Scholar
  13. 13.
    Bright JA, Taylor D, Curran JM, Buckleton JS (2013) Developing allelic and stutter peak height models for a continuous method of DNA interpretation. Forensic Sci Int Genet 7(2):296–304CrossRefPubMedGoogle Scholar
  14. 14.
    Criteria for the human fluorescent STR multiplex PCR reagent. GA/T 815–2009. China. Available from: http://www.bjstandard.com/standard/168830.html

Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Shuanglin Li
    • 1
  • Ling Chen
    • 2
  • Yanfang Wang
    • 3
  • Quyi Xu
    • 1
  • Hong Liu
    • 1
  • Yue Li
    • 1
  • Chao Liu
    • 1
  1. 1.Guangzhou Forensic Science InstituteGuangdong Province Key Laboratory of Forensic GeneticsGuangzhouChina
  2. 2.Institute of Forensic MedicineSouthern Medical UniversityGuangzhouChina
  3. 3.AGCU ScienTch IncorporationWuxiChina

Personalised recommendations