Forensic Science International: Genetics xxx (2011) xxx–xxx

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Letter to the Editor

Contents lists available at ScienceDirect

Forensic Science International: Genetics

jou r nal h o mep ag e: w ww .e lsev ier . co m / loc ate / fs ig

Allele frequencies of the new European Standard Set (ESS) lociplus SE33 locus in a population from the Republic of Macedonia

Dear Editor,

The highly polymorphic short tandem repeat (STR) loci in thehuman genome are widely used for forensic and paternity testingas well as for population genetic studies. Introducing set of STR locias observed markers made marvels progress in this field of forensicscience [1,2]. Nowadays, in response to the ENFSI and EDNAPgroups’ call for new STR multiplexes for Europe, Promegadeveloped a suite of four new DNA profiling kits. One of them isPowerPlex ESI 17 that we have used in this study.

In our previous population studies of Macedonian humanpopulation, we have used 15 STR loci included in the AmpFl-STR1Identifiler1 [3] and 17 Y-chromosomal short tandem repeatsloci incorporated in the AmpFistr1YfilerTM PCR Amplification Kit(data submitted to the FSI Genetics). All obtained results wereincluded in Macedonian referent database. In order of futuredevelopment of this database we have decide to analyze 6additional new STR loci (D22S1045, D1S1656, D10S1248, D2S441,D12S391, and SE33). Some of these loci were described even 15years ago [4,5].

This study is based on an observation of representative sampleof Macedonia human population. A total of 205 unrelatedindividuals of which 105 males and 100 females, from differentregions of Macedonia, participated in this study. All testedindividuals have been voluntary donors appropriately informedabout the research. Qiagen MicroTM Tissue Kit was used for DNAextraction from whole blood, Quantifiler1 assay for quantification

Table 1The distribution of allele frequencies and calculated forensic parameters for the Maced

Allele D1S1656 D2S441 D10S1248

2n 410 410 410

1 – – –

6.3 – – –

9 – 0.0024 –

10 – 0.1268 –

11 0.1146 0.3365 –

11.3 – 0.0951 –

12 0.1683 0.0415 0.0268

12.1 – 0.0039 –

12.3 – 0.0024 –

13 0.0683 0.0390 0.2341

13.2 – – –

14 0.0780 0.3073 0.3293

14.2 – – –

15 0.1244 0.0439 0.1780

15.3 0.0463 – –

16 0.1366 0.0024 0.1829

16.1 – – –

16.3 0.0390 – –

17 0.0341 – 0.0439

Please cite this article in press as: Z. Jakovski, et al., Allele frequenciespopulation from the Republic of Macedonia, Book review. Forensic

1872-4973/$ – see front matter � 2011 Elsevier Ireland Ltd. All rights reserved.

doi:10.1016/j.fsigen.2011.07.004

and PowerPlex ESI 171 System for amplification and detection(D18S51, D21S11, TH01, D3S1358, Amelogenin, D16S539,D2S1338, D1S1656, D10S1248, FGA, D8S1179, vWA, D22S1045,D19S433, D12S391, D2S441 and SE33). The total volume of eachreaction was 12.5 ml. The PCR amplifications have been carried outin ABI 9700 Thermal Cycler according to the manufacturer’srecommendations. Electrophoresis of the amplification productswas performed on an ABI 310 PRISM. Numerical allele designationsof the profiles were obtained by processing with Gene Mapper1

IDv 3.2 Software. Deviation from Hardy–Weinberg equilibrium,observed and expected heterozygosity, power of discriminationand power of exclusion were calculated. Also, we have comparedMacedonian data with data obtained from other neighboringpopulations with available data for observed loci. Deviation fromHardy–Weinberg equilibrium [6], observed and expected hetero-zygosity [7], exact test of population differentiation [8] andpairwise Fst [9] were calculated within Powermarker v3.25 [10],since power of discrimination and power of exclusion withinMicrosoft1 Excel workbook template—PowerStats [11].

Allele frequencies and all calculated forensic parameters areshown in Supplementary Table 1. All data are available on http://www.ingeb.ba/rebida/ESI17/macedonia.html.

No statistically significant deviation (p < 0.05) from Hardy-Weinberg equilibrium was found except for D1S1656 and SE33 loci(Table 1). Heterozygosity excess has been detected for SE33 locus.Allele frequencies of observed six loci within Macedonianpopulation have been compared with data obtained fromgeographically neighboring Southeast European populations suchas Serbia and Bosnia Herzegovina (our unpublished data). AfterBonferroni’s correction, statistical significance was considered as

onian population over 5 new ESS loci and SE33 locus (n = 205).

D12S391 SE33 D22S1045

410 410 410

– – –

– 0.0024 –

– – –

– – 0.0024

– – 0.1366

– – –

0.0049 0.0024 0.0122

– – –

– 0.0254 –

– 0.0122 –

– 0.0073 –

0.0024 0.0098 0.0537

– 0.0073 –

0.0512 0.0463 0.3634

– – –

0.0317 0.0366 0.3220

– 0.0024 –

– – –

0.1000 0.0659 0.0951

of the new European Standard Set (ESS) loci plus SE33 locus in aSci. Int. Genet. (2011), doi:10.1016/j.fsigen.2011.07.004

Table 1 (Continued )

Allele D1S1656 D2S441 D10S1248 D12S391 SE33 D22S1045

2n 410 410 410 410 410 410

17.2 – – – – 0.0049 –

17.3 0.0902 – – 0.0024 – –

18 0.0146 – 0.0024 0.1732 0.0756 0.0073

19 0.0024 – – 0.1171 0.1098 0.0073

19.2 – – – – 0.0049 –

19.3 0.0098 – – 0.0049 – –

20 – – – 0.0902 0.0537 –

20.2 – – – – 0.0122 –

20.3 – – – 0.0024 – –

21 – – – 0.1317 0.0122 –

21.2 – – – – 0.0122 –

22 – – – 0.1146 0.0073 –

22.2 – – – – 0.0366 –

23 – – – 0.0707 – –

24 – – 0.0024 0.0463 0.0024 –

24.1 – – – – 0.0024 –

24.2 – – – – 0.0390 –

25 – – – 0.0293 0.0024 –

25.2 – – – – 0.0244 –

26 – – – 0.0024 – –

27 – – – 0.0024 0.0024 –

27.2 – – – – 0.0756 –

28 – – – – 0.0024 –

28.2 – – – – 0.0512 –

29 – – – – 0.0024 –

29.2 – – – – 0.0610 –

30.2 – – – – 0.0488 –

31.2 – – – – 0.0146 –

32.2 – – – – 0.0220 –

33 – – – – 0.0024 –

33.2 – – – – 0.0073 –

34.2 – – – – 0.0024 –

35 – – – – 0.0073 –

36 – – – – 0.0024 –

37.2 – – – – 0.0024 –

PD 0.973 0.929 0.902 0.976 0.989 0.878

PE 0.791 0.506 0.599 0.721 0.810 0.579

P 0.0223 0.1076 0.2502 0.0582 0.0029 0.3163

H(ex) 0.8950 0.7636 0.7689 0.8955 0.9473 0.7334

H(ob) 0.8976 0.6976 0.8000 0.8634 0.9073 0.7854

PD, power of discrimination; PE, power of exclusion; P, Hardy–Weinberg equilibrium, exact test; H(ob), observed heterozygosity; H(ex), expected heterozygosity.

Letter to the Editor / Forensic Science International: Genetics xxx (2011) xxx–xxx2

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p < 0.015. Statistically significant differences of allele frequencieswere noticed for D12S391 locus (p = 0.0019) between Macedonianand Serbian population, as well as between Macedonian andBosnia Herzegovina population (p = 0.0005). Pairwise Fst, calcu-lated according to the results observed over these 6 loci, shows nosignificant genetic difference among Macedonian and Serbianpopulation (0.14%), as well as among Macedonian and BosniaHerzegovina population (0.15%).

According to our preliminary results, no disconcordancebetween PowerPlex ESI 17 and Identifiler or PowerPlex 16 wasrecorded. Detailed concordance study, which is going to includealmost thousand samples from this part of Europe, is ongoing.

In the course of the elaboration of this population study, weadhered to the new guidelines published in this journal, whereinwe would like to point out that we accept the requirementspertaining to the publication of this type of papers [12], as well aISFG recommendations [13]. Also we follow recommendations ofthe European Council resolution (9192/01) calls upon EU countriesto use the European Standard Set (ESS) as a minimum to enableinternational comparison of DNA profiles as well as the paper forthe evolution of DNA databases and recommendations [14].

Acknowledgment

We would like to thank to the Promega Corporation forsupporting this study.

Please cite this article in press as: Z. Jakovski, et al., Allele frequenciespopulation from the Republic of Macedonia, Book review. Forensic

References

[1] P. Gill, C.P. Kimpton, A. Urquhart, et al., Automated short tandem repeat (STR)analysis in forensic casework—a strategy for the future, Electrophoresis 16 (1995)1543–1552.

[2] J.A. Thomson, V. Pilotti, P. Stevens, K.L. Ayres, P.G. Debenham, Validation of shorttandem repeat analysis for the investigation of cases of disputed paternity,Forensic Sci. Int. 100 (1998) 1–16.

[3] Z. Jakovski, K. Nikolova, I. Furac, M. Masic, B. Janeska, M. Kubat, Allele frequenciesfor 15 STR loci in a population from the Republic of Macedonia, Int. J. Legal Med.120 (2006) 53–55.

[4] M.V. Lareu, C. Pestoni, M. Schurenkamp, S. Rand, B. Brinkmann, A. Carracedo, Ahighly variable STR at the D12S391 locus, Int. J. Legal Med. 109 (3) (1996) 134–138.

[5] M.V. Lareu, S. Barral, A. Salas, C. Pestoni, A. Carracedo, Sequence variation of ahypervariable short tandem repeat at the D1S1656 locus, Int. J. Legal Med. 111 (5)(1998) 244–247.

[6] S.W. Guo, E.A. Thompson, Performing the exact test of Hardy–Weinberg propor-tion for multiple alleles, Biometrics 48 (1992) 361–372.

[7] M. Nei, Molecular Evolutionary Genetics, Columbia University Press, New York,1987.

[8] M. Raymond, F. Rousset, An exact test for population differentiation, Evolution 49(1995) 1280–1283.

[9] B.S. Weir, Genetic Data Analysis II: Methods for Discrete Population Genetic Data,Sinauer Assoc., Inc., Sunderland, MA, 1996.

[10] L. Kejun, V.M. Spencer, PowerMarker: an integrated analysis environment forgenetic marker analysis, Bioinformatics 21 (2005) 2128–2129.

[11] A. Tereba, Tools for analysis of population statistics, Profiles DNA 3 (1999) 14–16.[12] A. Carracedo, J.M. Butler, L. Gusmao, W. Parson, L. Roewer, P.M. Schneider,

Publication of population data for forensic purposes, Forensic Sci. Int. Genet. 4(2010) 145–147.

[13] B. Olaisen, W. Bar, B. Brinkmann, B. Budowle, A. Carracedo, P. Gill, P. Lincoln, W.R.Mayr, S. Vox Sang Rand, DNA recommendations 1997 of the International Societyfor Forensic Genetics, Vox Sang. 74 (1) (1998) 61–63.

of the new European Standard Set (ESS) loci plus SE33 locus in aSci. Int. Genet. (2011), doi:10.1016/j.fsigen.2011.07.004

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[14] P. Gill, L. Fereday, N. Morling, P.M. Schneider, The evolution of DNA databases—recommendations for new European STR loci, Forensic Sci. Int. 156 (2006) 242–244.

Zlatko Jakovski*

Ksenija NikolovaRenata Jankova-Ajanovska

Biljana JaneskaInstitute of Forensic Medicine and Criminology,

School of Medicine, University ‘‘Ss. Cyril and Methodious’’,

Skopje, Macedonia

Naris PojskicInstitute for Genetic Engineering and Biotechnology,

Sarajevo, Bosnia and Herzegovina

Please cite this article in press as: Z. Jakovski, et al., Allele frequenciespopulation from the Republic of Macedonia, Book review. Forensic

Damir Marjanovicab

aInstitute for Genetic Engineering and Biotechnology,

Sarajevo, Bosnia and HerzegovinabGenos, Planinska 1, 10000 Zagreb, Croatia

*Corresponding author. Tel.: +389 2 3 177 044;fax: +389 2 1 178 831

E-mail address: zlatedr@yahoo.com (Z. Jakovski).

24 December 2010

of the new European Standard Set (ESS) loci plus SE33 locus in aSci. Int. Genet. (2011), doi:10.1016/j.fsigen.2011.07.004