Molecular Biology Reports

, Volume 44, Issue 1, pp 183–189 | Cite as

Analysis of 12 X-STR loci in the population of south Croatia

  • Gordan Mršić
  • Petar Ozretić
  • Josip Crnjac
  • Siniša Merkaš
  • Ivana Račić
  • Sara Rožić
  • Viktorija Sukser
  • Maja Popović
  • Marina Korolija
Original Article


The aim of the study was to assess forensic pertinence of 12 short tandem repeats (STRs) on X-chromosome in south Croatia population. Investigator® Argus X-12 kit was used to co-amplify 12 STR loci belonging to four linkage groups (LGs) on X-chromosome in 99 male and 98 female DNA samples of unrelated donors. PCR products were analyzed by capillary electrophoresis. Population genetic and forensic parameters were calculated by the Arlequin and POPTREE2 software, and an on-line tool available at Hardy–Weinberg equilibrium was confirmed for all X-STR markers in female samples. Biallelic patterns at DXS10079 locus were detected in four male samples. Polymorphism information content for the most (DXS10135) and the least (DXS8378) informative markers was 0.9212 and 0.6347, respectively. In both male and female samples, combined power of discrimination exceeded 0.999999999. As confirmed by linkage disequilibrium test, significant association of marker pair DXS10074-DXS10079 (P = 0.0004) within LG2 and marker pair DXS10101-DXS10103 (P = 0.0003) within LG3 was found only in male samples. Number of observed haplotypes in our sample pool amounted 3.01, 7.53, 5 and 3.25% of the number of possible haplotypes for LG1, LG2, LG3 and LG4, respectively. According to haplotype diversity value of 0.9981, LG1 was the most informative. In comparison of south Croatia with 26 world populations, pair-wise \({{\text{F}}_{\text{ST}}}^{\text{*}}\) values increase in parallel with geographical distance. Overall statistical assessment confirmed suitability of Investigator® Argus X-12 kit for forensic casework in both identification and familial testing in the population of south Croatia.


X-STR Investigator® Argus X-12 Population study South Croatia Forensics 



We are particularly grateful for having had the opportunity to collaborate with Branka Gršković, our late colleague, who had generously dedicated her time and great merit in making this research possible.


This study was funded by the Ministry of the Interior of Croatia.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11033_2017_4096_MOESM1_ESM.pdf (208 kb)
Supplementary material 1 (PDF 208 KB)
11033_2017_4096_MOESM2_ESM.xlsx (35 kb)
Supplementary material 2 (XLSX 35 KB)


  1. 1.
    Szibor R, Hering S, Edelmann J (2006) A new web site compiling forensic chromosome X research is now online. Int J Legal Med 120(4):252–254. doi: 10.1007/s00414-005-0029-y CrossRefPubMedGoogle Scholar
  2. 2.
    Diegoli TM (2014) Forensic application of X chromosome STRs. In: Primorac D, Schanfield M (eds) Forensic DNA applications: an interdisciplinary perspective, vol 1. CRC Press, New York, pp 135–171CrossRefGoogle Scholar
  3. 3.
    Branicki W, Wolanska-Nowak P, Parys-Proszek A, Kupiec T (2008) Application of the Mentype Argus X-8 kit to forensic casework. Probl Forensic Sci 73:12Google Scholar
  4. 4.
    Pinto N, Gusmao L, Amorim A (2011) X-chromosome markers in kinship testing: a generalisation of the IBD approach identifying situations where their contribution is crucial. Forensic Sci Int 5 (1):27–32. doi: 10.1016/j.fsigen.2010.01.011 CrossRefGoogle Scholar
  5. 5.
    Diegoli TM (2015) Forensic typing of short tandem repeat markers on the X and Y chromosomes. Forensic Sci Int 18:140–151. doi: 10.1016/j.fsigen.2015.03.013 CrossRefGoogle Scholar
  6. 6.
    Tillmar AO, Mostad P, Egeland T, Lindbolm B, Holmlund G, Montelius K (2008) Analysis of linkage and linkage disequilibrium for eight X-STR markers. Forensic Science Int 3 (1):37–41. doi: 10.1016/j.fsigen.2008.09.006 CrossRefGoogle Scholar
  7. 7.
    Gomes C, Magalhaes M, Alves C, Amorim A, Pinto N, Gusmao L (2012) Comparative evaluation of alternative batteries of genetic markers to complement autosomal STRs in kinship investigations: autosomal indels vs. X-chromosome STRs. Int J Legal Med 126(6):917–921. doi: 10.1007/s00414-012-0768-5 CrossRefPubMedGoogle Scholar
  8. 8.
    Szibor R, Krawczak M, Hering S, Edelmann J, Kuhlisch E, Krause D (2003) Use of X-linked markers for forensic purposes. Int J Legal Med 117(2):67–74. doi: 10.1007/s00414-002-0352-5 PubMedGoogle Scholar
  9. 9.
    Walsh PS, Metzger DA, Higuchi R (1991) Chelex-100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques 10(4):506–513PubMedGoogle Scholar
  10. 10.
    Excoffier L, Lischer HEL (2010) Arlequin suite ver 3.5: a new series of programs to perform population genetics analyses under Linux and Windows. Mol Ecol Resour 10(3):564–567. doi: 10.1111/j.1755-0998.2010.02847.x CrossRefPubMedGoogle Scholar
  11. 11.
    Latter BD (1972) Selection in finite populations with multiple alleles.III. Genetic divergence with centripetal selection and mutation. Genetics 70(3):475–490PubMedPubMedCentralGoogle Scholar
  12. 12.
    Takezaki N, Nei M, Tamura K (2010) POPTREE2: software for constructing population trees from allele frequency data and computing other population statistics with Windows interface. Mol Biol Evol 27(4):747–752. doi: 10.1093/molbev/msp312 CrossRefPubMedGoogle Scholar
  13. 13.
    Poulsen L, Tomas C, Drobnic K, Ivanova V, Mogensen HS, Kondili A, Miniati P, Bunokiene D, Jankauskiene J, Pereira V, Morling N (2016) NGMSElect and Investigator((R)) Argus X-12 analysis in population samples from Albania, Iraq, Lithuania, Slovenia, and Turkey. Forensic Sci Int 22:110–112. doi: 10.1016/j.fsigen.2016.02.004 CrossRefGoogle Scholar
  14. 14.
    Bekada A, Benhamamouch S, Boudjema A, Fodil M, Menegon S, Torre C, Robino C (2010) Analysis of 21 X-chromosomal STRs in an Algerian population sample. Int J Legal Med 124(4):287–294. doi: 10.1007/s00414-009-0397-9 CrossRefPubMedGoogle Scholar
  15. 15.
    Rebala K, Kotova SA, Rybakova VI, Zabauskaya TV, Shyla AA, Spivak AA, Tsybovsky IS, Szczerkowska Z (2015) Variation of X-chromosomal microsatellites in Belarus within the context of their genetic diversity in Europe. Forensic Sci Int 16:105–111. doi: 10.1016/j.fsigen.2014.12.011 CrossRefGoogle Scholar
  16. 16.
    Bentayebi K, Picornell A, Bouabdeallah M, Castro JA, Aboukhalid R, Squalli D, Misericordia M, Amzazi S (2012) Genetic diversity of 12 X-chromosomal short tandem repeats in the Moroccan population. Forensic Sci Int 6(1):e48–e49. doi: 10.1016/j.fsigen.2011.03.008 CrossRefGoogle Scholar
  17. 17.
    Crnjac J, Ozretić P, Merkaš S, Ratko M, Lozančić M, Rožić S, Špoljarić D, Korolija M, Popović M, Mršić G (2016) Analysis of 12 X-chromosomal markers in the population of central Croatia. Legal Med 21:77–84. doi: 10.1016/j.legalmed.2016.07.001 CrossRefPubMedGoogle Scholar
  18. 18.
    Zidkova A, Capek P, Horinek A, Coufalova P (2014) Investigator (R) Argus X-12 study on the population of Czech Republic: comparison of linked and unlinked X-STRs for kinship analysis. Electrophoresis 35(14):1989–1992. doi: 10.1002/elps.201400046 CrossRefPubMedGoogle Scholar
  19. 19.
    Poulsen L, Farzad MS, Borsting C, Tomas C, Pereira V, Morling N (2015) Population and forensic data for three sets of forensic genetic markers in four ethnic groups from Iran: Persians, Lurs, Kurds and Azeris. Forensic Sci Int 17:43–46. doi: 10.1016/j.fsigen.2015.03.010 CrossRefGoogle Scholar
  20. 20.
    Tomas C, Pereira V, Morling N (2012) Analysis of 12 X-STRs in Greenlanders, Danes and Somalis using Argus X-12. Int J Legal Med 126(1):121–128. doi: 10.1007/s00414-011-0609-y CrossRefPubMedGoogle Scholar
  21. 21.
    Edelmann J, Lutz-Bonengel S, Naue J, Hering S (2012) X-chromosomal haplotype frequencies of four linkage groups using the investigator Argus X-12 kit. Forensic Sci Int 6 (1):E24–E34. doi: 10.1016/j.fsigen.2011.01.001 CrossRefGoogle Scholar
  22. 22.
    Tomas C, Skitsa I, Steinmeier E, Poulsen L, Ampati A, Borsting C, Morling N (2015) Results for five sets of forensic genetic markers studied in a Greek population sample. Forensic Sci Int 16:132–137. doi: 10.1016/j.fsigen.2015.01.001 CrossRefGoogle Scholar
  23. 23.
    Horvath G, Zalan A, Kis Z, Pamjav H (2012) A genetic study of 12 X-STR loci in the Hungarian population. Forensic Sci Int 6 (1):E46–E47. doi: 10.1016/j.fsigen.2011.03.007 CrossRefGoogle Scholar
  24. 24.
    Ferragut JF, Bentayebi K, Castro JA, Ramon C, Picornell A (2015) Genetic analysis of 12 X-chromosome STRs in Western Mediterranean populations. Int J Legal Med 129(2):253–255. doi: 10.1007/s00414-014-1071-4 CrossRefPubMedGoogle Scholar
  25. 25.
    Bini C, Riccardi LN, Ceccardi S, Carano F, Sarno S, Luiselli D, Pelotti S (2015) Expanding X-chromosomal forensic haplotype frequencies database: Italian population data of four linkage groups. Forensic Sci Int 15:127–130. doi: 10.1016/j.fsigen.2014.11.008 CrossRefGoogle Scholar
  26. 26.
    Shrivastava P, Jain T, Gupta U, Trivedi VB (2015) Genetic polymorphism study on 12 X STR loci of investigator Argus X STR kit in Bhil tribal population of Madhya Pradesh, India. Legal Med 17 (3):214–217. doi: 10.1016/j.legalmed.2014.11.004 CrossRefPubMedGoogle Scholar
  27. 27.
    Caine L, Costa S, Pinheiro MF (2013) Population data of 12 X-STR loci in a North of Portugal sample. Int J Legal Med 127(1):63–64. doi: 10.1007/s00414-012-0672-z CrossRefPubMedGoogle Scholar
  28. 28.
    Tillmar AO (2012) Population genetic analysis of 12 X-STRs in Swedish population. Forensic Sci Int 6(2):e80–e81. doi: 10.1016/j.fsigen.2011.07.008 CrossRefGoogle Scholar
  29. 29.
    Uchigasaki S, Tie J, Takahashi D (2013) Genetic analysis of twelve X-chromosomal STRs in Japanese and Chinese populations. Mol Biol Rep 40(4):3193–3196. doi: 10.1007/s11033-012-2394-1 CrossRefPubMedGoogle Scholar
  30. 30.
    Costa HA, Morais P, Da Silva C, Matos S, Santos RM, Espinheira R, Santos JC, Amorim A (2014) X-chromosome STR markers data in a Cabo Verde immigrant population of Lisboa. Mol Biol Rep 41(4):2559–2569. doi: 10.1007/s11033-014-3114-9 CrossRefGoogle Scholar
  31. 31.
    Hundertmark T, Hering S, Edelmann J, Augustin C, Plate I, Szibor R (2008) The STR cluster DXS10148-DXS8378-DXS10135 provides a powerful tool for X-chromosomal haplotyping at Xp22. Int J Legal Med 122(6):489–492. doi: 10.1007/s00414-008-0277-8 CrossRefPubMedGoogle Scholar
  32. 32.
    Sim JE, Lee HY, Yang WI, Shin KJ (2010) Population genetic study of four closely-linked X-STR trios in Koreans. Mol Biol Rep 37(1):333–337. doi: 10.1007/s11033-009-9733-x CrossRefPubMedGoogle Scholar
  33. 33.
    Czarnogorska M, Sanak M, Piniewska D, Polanska N, Stawowiak A, Opolska-Bogusz B (2010) [Identification of rare genetic variants at DXS10074, DXS10079, DXS10146 and DXS10148 loci of investigator argus X-12 multiplex in the south Polish population]. Arch Med Sadowej Kryminol 60(4):235–242PubMedGoogle Scholar
  34. 34.
    Zhang S, Zhao S, Zhu R, Li C (2012) Genetic polymorphisms of 12 X-STR for forensic purposes in Shanghai Han population from China. Mol Biol Rep 39(5):5705–5707. doi: 10.1007/s11033-011-1379-9 CrossRefPubMedGoogle Scholar
  35. 35.
    Tri-Allelic Patterns (2015) Accessed 16 Feb 2016
  36. 36.
    Clayton TM, Guest JL, Urquhart AJ, Gill PD (2004) A genetic basis for anomalous band patterns encountered during DNA STR profiling. J Forensic Sci 49(6):1207–1214CrossRefPubMedGoogle Scholar
  37. 37.
    Rolf B, Wiegand P, Brinkmann B (2002) Somatic mutations at STR loci—a reason for three-allele pattern and mosaicism. Forensic Sci Int 126(3):200–202CrossRefPubMedGoogle Scholar
  38. 38.
    Grskovic B, Zidkova A, Stenzl V, Popovic M, Primorac D, Mrsic G (2013) Analysis of 8 X-chromosomal markers in the population of central Croatia. Croat Med J 54(3):238–247. doi: 10.3325/cmj.2013.54.238 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  • Gordan Mršić
    • 1
    • 2
  • Petar Ozretić
    • 3
  • Josip Crnjac
    • 4
  • Siniša Merkaš
    • 1
    • 2
  • Ivana Račić
    • 1
    • 2
  • Sara Rožić
    • 1
    • 2
  • Viktorija Sukser
    • 1
    • 2
  • Maja Popović
    • 5
  • Marina Korolija
    • 1
    • 2
  1. 1.Forensic Science Centre “Ivan Vučetić”ZagrebCroatia
  2. 2.Forensic Science OfficeUniversity of ZagrebZagrebCroatia
  3. 3.Ruđer Bošković InstituteZagrebCroatia
  4. 4.University Department for Forensic SciencesUniversity of SplitSplitCroatia
  5. 5.Faculty of Veterinary MedicineUniversity of ZagrebZagrebCroatia

Personalised recommendations