Forensic Science International: Genetics 5 (2011) e108e111

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Forensic Science International: Genetics

journal homepage: www.elsevier.com/locate/fsig

Forensic Population Genetics Short Communication

Genetic data for 17 Y-chromosomal STR loci in Macedonians in the

Republic of Macedonia
Zlatko Jakovski a,*, Ksenija Nikolova a, Renata Jankova-Ajanovska a, Damir Marjanovic b,c,
Naris Pojskic b, Biljana Janeska a
a
Institute of Forensic Medicine and Criminology, School of Medicine, University Ss. Cyril and Methodious, Skopje, Macedonia
b
Institute for Genetic Engineering and Biotechnology, Sarajevo, Bosnia and Herzegovina
c
Genos, Planinska 1, 10000 Zagreb, Croatia

A R T I C L E I N F O A B S T R A C T

Article history: The population data were obtained for the 16 Y chromosomal STR loci included in the AmpFistr1YlerTM
Received 14 July 2010 PCR Amplication Kit (DYS456, DYS389I, DYS390, DYS389II, DYS458, DYS19, DYS385 a/b, DYS393,
Received in revised form 10 November 2010 DYS391, DYS439, DYS635, DYS392, Y-GATA H4, DYS437, DYS438, DYS448) in a sample of 262 unrelated
Accepted 2 April 2011
men from the Republic of Macedonia.
2011 Elsevier Ireland Ltd. All rights reserved.
Keywords:
Y chromosomal short tandem repeats (STR)
Macedonian population data
AmpFistr1YlerTM PCR Amplication Kit

Populations: This population study was carried out to establish
a forensic database on the Macedonian population. The population
data were obtained for the 16 Y chromosomal STR loci included in
the AmpFistr1YlerTM PCR Amplication Kit (Applied Biosysetm)
(DYS456, DYS389I, DYS390, DYS389II, DYS458, DYS19, DYS385 a/b,
DYS393, DYS391, DYS439, DYS635, DYS392, Y-GATA H4, DYS437,
DYS438, DYS448). Similar population databases were found for
most countries in the area [18] (Fig. 1). The DNA samples
originate from 262 unrelated males. In view of the fact that the
Republic of Macedonia is a multi-national country with inhabi-
tants of various ethnic origins and religions, it would be worth
noting that the foregoing males are Orthodox Christians of
Macedonian ethnicity from different geographical regions in the
Republic of Macedonia.
DNA extraction: Whole blood samples were collected on FTA
classic cards (Whatman) and stored at room temperature. DNA
from whole blood was extracted with QiaAmp DNA MiniKit
(QIAGEN Gmbh, Hilden Germany).
PCR: One multiplex PCR reaction was carried out for each DNA
sample using the AmpFistr1YlerTM PCR Amplication Kit
(Applied Biosysetm) according to the suppliers protocols. Ampli-
cation was carried out in a 9700 Thermal Cycler (Applied
Biosystems).
Typing: For electrophoresis, 1.5 ml of the PCR product was
combined with 12 ml of formamide and 0.5 ml of GeneScan 500 LIZ
* Corresponding author. Tel.: +389 2 3 177 0 44; fax: +389 2 1 178 831.
E-mail address: zlatedr@yahoo.com (Z. Jakovski).
size standard. Detection of PCR products and genotyping were
carried out on the ABI PRISM 310 Genetic Analyser (Applied
Biosystems) using the data collection software and GeneMapper ID
v3.2 analysis software (Applied Biosystems). The allele nomencla-
ture is according to the Y-STR Haplotype Reference Database, YHRD
(Y-STR Haplotype Reference Database, http://www.yhrd.org).
Data analysis: An estimation of allele and haplotype frequency
distribution, haplotype and gene diversity was done using Neis
statistics [9]. An exact test of sample differentiation [10] based on
haplotype frequencies have been calculated in order to estimate a
possible difference among the Macedonian and the other observed
control populations. The considered signicance level after Bonfer-
ronis correction was p < 0.01. In order to determine the Macedonian
population Y_STR genetic relationships with the populations in the
neighboring regions (Bosnia and Herzegovina, Bulgaria, Croatia,
Kosovo, Serbia), share allele genetic distance [11] has been applied.
Also neighbor-joining dendrogram [12] based on result of genetic
distance analysis has been constructed. Consensus tree has been
constructed upon 1000 bootstraps. Principal components (PC)
analysis was performed on allele frequencies by using PAST software
[13]. Since only 7 same Y-STR loci were available from previous
studies for all included populations from neighboring regions,
DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, loci
have been considered during the genetic distance analysis.
The above mentioned calculations were made using Arlequin
version 3.11 [14], PowerMaker version 3.32 [15] and MEGA4 [16]
software.
Access to the data: Available upon request by email to:
zlatedr@yahoo.com.

1872-4973/$ see front matter 2011 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.fsigen.2011.04.005
[()TD$FIG] Z. Jakovski et al. / Forensic Science International: Genetics 5 (2011) e108e111 e109

Fig. 1. Geographical location of Republic of Macedonia (red label) and other neighboring countries which population data were used (yellow label).

Results: Total of 243 different Y-STR haplotypes from 262 and haplotype diversity within the samples of the Macedonian
samples were detected: 227 of them were unique, 16 haplotypes population are presented in Tables 1 and 2. Major allele
appeared twice and just one three times. An estimation of frequency and gene diversity across loci are presented in
allele frequencies distribution, the most frequent haplotypes Table 3.

Table 1
Allele frequency distribution of observed Y STR loci in the Macedonian population.

Marker Allele Count Freq % Marker Allele Count Freq %

DYS19 13 73 0.2786 27.9 DYS438 10 156 0.5954 59.5

14 54 0.2061 20.6 11 40 0.1527 15.3
15 64 0.2443 24.4 12 25 0.0954 9.5
16 56 0.2137 21.4 13 1 0.0038 0.4
17 15 0.0573 5.7 8 1 0.0038 0.4
DYS389I 11 1 0.0038 0.4 9 39 0.1489 14.9
12 43 0.1641 16.4 DYS439 10 24 0.0916 9.2
13 180 0.6870 68.7 11 68 0.2595 26.0
14 37 0.1412 14.1 12 114 0.4351 43.5
9 1 0.0038 0.4 13 44 0.1679 16.8
DYS389II 27 1 0.0038 0.4 14 11 0.0420 4.2
28 31 0.1183 11.8 15 1 0.0038 0.4
29 47 0.1794 17.9 DYS437 14 148 0.5649 56.5
30 103 0.3931 39.3 14.1 1 0.0038 0.4
31 61 0.2328 23.3 15 80 0.3053 30.5
32 16 0.0611 6.1 16 33 0.1260 12.6
33 3 0.0115 1.1 DYS448 16 2 0.0076 0.8
DYS390 21 2 0.0076 0.8 18 7 0.0267 2.7
22 15 0.0573 5.7 19 77 0.2939 29.4
23 52 0.1985 19.8 20 146 0.5573 55.7
24 156 0.5954 59.5 21 28 0.1069 10.7
25 36 0.1374 13.7 22 2 0.0076 0.8
26 1 0.0038 0.4 DYS456 12 1 0.0038 0.4
DYS391 10 162 0.6183 61.8 13 16 0.0611 6.1
11 86 0.3282 32.8 14 19 0.0725 7.3
12 2 0.0076 0.8 15 109 0.4160 41.6
9 12 0.0458 4.6 15.3 1 0.0038 0.4
DYS392 10 2 0.0076 0.8 16 74 0.2824 28.2
11 222 0.8473 84.7 16.1 1 0.0038 0.4
12 11 0.0420 4.2 17 32 0.1221 12.2
13 25 0.0954 9.5 18 8 0.0305 3.1
14 1 0.0038 0.4 21 1 0.0038 0.4
16 1 0.0038 0.4 DYS458 13 1 0.0038 0.4
DYS393 10 1 0.0038 0.4 14 14 0.0534 5.3
12 60 0.2290 22.9 15 79 0.3015 30.2
13 185 0.7061 70.6 16 71 0.2710 27.1
14 13 0.0496 5.0 17 67 0.2557 25.6
15 3 0.0115 1.1 17.2 6 0.0229 2.3
DYS385 (a and b) 10 3 0.0057 0.6 18 14 0.0534 5.3
11 62 0.1183 11.8 19 4 0.0153 1.5
12 9 0.0172 1.7 20 5 0.0191 1.9
13 43 0.0821 8.2 21 1 0.0038 0.4
14 118 0.2252 22.5 DYS635 20 11 0.0420 4.2
15 70 0.1336 13.4 21 58 0.2214 22.1
15.3 2 0.0038 0.4 22 64 0.2443 24.4
16 88 0.1679 16.8 23 109 0.4160 41.6
16.3 1 0.0019 0.2 24 17 0.0649 6.5
17 50 0.0954 9.5 25 2 0.0076 0.8
18 61 0.1164 11.6 26 1 0.0038 0.4
19 11 0.0210 2.1 GATA-H4 10 3 0.0115 1.1
20 4 0.0076 0.8 11 117 0.4466 44.7
21 2 0.0038 0.4 12 112 0.4275 42.7
13 25 0.0954 9.5
14 1 0.0038 0.4
9 4 0.0153 1.5
e110 Z. Jakovski et al. / Forensic Science International: Genetics 5 (2011) e108e111

Table 2 Table 3
Haplotypes that have been detected more then once in the Macedonian population Major allele frequency and gene diversity for observed Y STR loci in examined
sample and observed haplotype diversity. sample of the Macedonian population.

Haplotypes Count Locus Major allele freq. Allele no. Gene diversity

17-13-31-25-12-11-13-13-15-10-13-15-20-15-17-21-11 2 DYS19 0.2786 5 0.7713

16-13-31-22-10-11-12-13-15-9-12-14-21-15-15-21-12 2 DYS389I 0.6870 5 0.4811
17-13-31-25-11-11-13-11-13-11-11-14-20-15-15-23-13 2 DYS389II 0.3931 7 0.7412
13-13-30-24-11-11-13-16-18-10-12-14-20-16-15-23-12 2 DYS390 0.5954 6 0.5839
16-13-31-24-11-11-13-14-15-10-13-15-19-15-17-23-11 2 DYS391 0.6183 4 0.5078
16-13-30-24-10-11-13-11-14-11-10-14-20-16-15-24-13 2 DYS392 0.8473 6 0.2711
14-12-28-24-10-11-12-14-17-9-12-16-18-13-15-22-11 2 DYS393 0.7061 5 0.4464
13-13-30-24-10-11-13-16-18-10-12-14-20-17-17-22-12 3 DYS385 a and b 0.2252 14 0.8590
16-14-31-25-10-11-13-11-14-11-11-14-20-16-16-23-12 2 DYS438 0.5954 6 0.5909
14-13-29-24-11-13-12-11-11-12-13-15-19-15-15-23-12 2 DYS439 0.4351 6 0.7049
15-12-28-23-9-11-12-13-16-9-14-14-21-16-14-22-12 2 DYS437 0.5649 4 0.5718
16-13-30-23-10-11-13-11-14-11-10-14-21-17-16-23-12 2 DYS448 0.5573 6 0.5908
15-13-30-25-10-11-13-11-14-11-11-14-20-18-16-23-12 2 DYS456 0.4160 10 0.7222
15-12-29-22-10-11-13-14-14-10-12-16-21-15-16-20-11 2 DYS458 0.3015 10 0.7634
14-13-30-24-10-13-13-11-11-12-12-14-19-15-17-23-12 2 DYS635 0.4160 7 0.7122
13-14-31-24-10-11-13-16-18-10-11-14-20-16-15-22-12 2 GATA-H4 0.4466 6 0.6084
15-13-31-24-10-11-13-17-18-10-12-14-20-17-16-23-12 2 Mean 0.5052 6.6875 0.6204
Haplotype diversity 1.0000  0.0003

Table 4
There are no statistical signicant differences in allele Share allele distance among observed populations based on 7 observed Y STR loci.

frequencies between the Macedonian and the other observed Population B&H Bulgaria Croatia Kosovo Macedonia Serbia
populations (p > 0.01).
B&H
The Macedonian population has the largest genetic distance Bulgaria 0.1579
against the Croatian population (0.2643), while the lowest against Croatia 0.2036 0.2687
Bulgarian population (0.0815) (Table 4). The neighbor-joining Kosovo 0.2321 0.1766 0.3291
Macedonia 0.1376 0.0815 0.2643 0.1543
consensus tree constructed based on the results of the genetic
Serbia 0.1155 0.1128 0.2160 0.2064 0.1157
distance analysis shows that the population of the Macedonian
ethnic origin has possible highest similarity with the Bulgarian
populations, and not such dissimilarity with the cluster which
consists Bosnian and Herzegovinian-Serbian-Croatian populations, percentage of unique haplotype (84.64%) is very high as well as
while the Kosovo population create a separate group (Fig. 2). haplotype diversity. Gene diversity shows variation over loci in
The rst component PCA reveals that Macedonian population is range 0.27110.8590 with overall average value 0.6204.
genetically closest to the Bulgarian and partially Serbian popula- The Balkan region includes human populations with high level
tion especially compared with Kosovo population. The second of Y STR diversity and with a quite similar genetic structure. It is
component PCA shows also closest relation of Macedonian and clear that the observed populations from central and eastern
Bulgarian populations. Croatian population is located at the other Balkan do not have a pronounced differentiation of Y STR
end of the dimension since Bosnia and Herzeogina population has population structure, except partially Kosovo population which
separated position in compared with Macedonia and Bulgaria shows some of Y STR specicity. As the obtained results indicate,
populations. Kosovo population clusters separately among studied genetic distance recorded between Macedonian population and
populations. Total variance of the data set was 62%, from which the ethnic Albanians from Kosovo is relatively high. This nding is
rst and second PCs represented, 36% and 26% respectively (Fig. 3). curious considering that Kosovo and Republic Macedonia share a
Conclusion: Previously published data for the Macedonian border. In addition, almost 25% of Republic Macedonia citizens
population which were announced 5 years ago were based on the constitute ethnic Albanians who share the origins with Albanian
observation of the 10 loci [1]. The main intention of this study was population in Kosovo. Therefore, the results of this study suggest
to expand published Y-STR genetic data for the Macedonian that forensic experts in Macedonia need to apply caution when
population with the information from the seven new loci, as well to estimating haplotype frequencies that are to be used in the
present some additional information about the Y-STR genetic quantitative assessment of matches in forensic and genealogical
relationships of the Macedonian population with the human casework. Ethnicity of the examined individuals needs to be taken
populations from the neighboring countries. into consideration and appropriate population database used in
According to the results of the comparative analysis, we can these estimations.
conclude that population differences between the six observed In the course of the elaboration of this population study, we
populations are expected. Macedonian population shows moder- adhered to the new guidelines published in this journal, wherein
ate Y-STR differentiation versus other Balkan populations. The we would like to point out that we accept the requirements
[()TD$FIG]
Kosovo
Bulgaria
68 Croatia
96
Bosnia and Herzegovina
92
Serbia
Macedonia

Fig. 2. Neighbor-joining consensus tree (1000 bootstraps) based on share allele genetic distance analyze estimated using the available data for 7 Y STR loci.
[()TD$FIG] Z. Jakovski et al. / Forensic Science International: Genetics 5 (2011) e108e111
Component 2
-2 -1.6 -1.2 -0.8 -0.4
Bosnia_and_Herzegovina 0.4
Croatia
Component 1
Fig. 3. PC analysis performed using allele frequencies of available data for 7 Y STR loci.
pertaining to the publication of this type of papers [17].
Furthermore, this paper is in line both with the original [18],
and the updated ISFG guidelines [19] of the DNA Commission of the
International Society of Forensic Genetics on the use of Y-STRs in
forensic analysis.
Quality control: GEDNAP (German DNA Proling Group) blind
trial system [20].
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