ALLELE FREQUENCIES DISTRIBUTION OF 5 STR SYSTEMS IN ÇUKUROVA REGION

<;ukurova

Yoresinde 5 STR Sistemi (HUMTHOl

,

HUMvWA, HU1v.1F13

B

,

HUMCSFIPO,

HUMD21S11)'nin Allel Frekans Dagllmu

Alper

B*, Se

rin A*,

S~hiirenkamp

M**

,

D

ag

H*

, Brinkmann

B**

*(:ukurova

Universitesi T

I

p

Fakii

l

tesi A

dl

i

TIP

Anabilim D

a

b,

Adana, TURKjYE

**WestfAlische-Wilhelms

Universitesi

,

Adli

TIP

Enstitiisii,

Mii

n

ster,

ALMANYA.

Ozet

B

u

~a!J§mada

9ukurova yoresinde ya§ayan

110 ki§i1ik

biT

popuJasyon orneginde

Hum

TH0

1

,

H

umvW

A,

HumF13B,

H

um

CSF

I

P

O

ve

HumD2

1

S

11

STR

lokuslannm allel

frekansl

a

n

saptanml§tII. Belirtilen

STR

lokuslarmm

bu yored

e

adii ama~lI

kimIiklendirme

ve akraballk

jJj

§kilerin

in

saptanmasmda

kullamlabilirJig

i

istatistiksel olarak degerlend

i

ril

-mi§ ve eJd

e ed

ilen

sonu~lar

farklI populasyon

~aII§malan

ile kar§lla§tmlml§tlr

.

Rastge

le

se~iIen

ve araIarmda akraballk ili§kisi bulunmayan sagbkii vericilerden alman

kan orneklerinden deneysel olarak olu§turuJan kan lekelerinde

Che

l

ex

ile

DNA

ekstraksi-yonun

u takiben

PCR, PAG

elektroforezi ve

giimii§

boyama yontemi ile analiz

yapllml§tlr.

9a!J§mada

7

THO

I alleli (6-

11),8

v

WA

alleli

(

1

3-2

0)

,5

F13B

alleli

(6-10),6

CSF

J

PO

alleli

(9

-

14)

ve

12

D21S11 alleli

(26-34.2)

tammJanml

§tlr

.

9alI§1lan

5

si

s

temin alleI frekans

dagllImlannm

H

ardy-

W

einberg

Dengesi

ile uyumlu

oldugu

sap

tanml§tI

r

.

C;u

kurova yor

e

si

i~in

hesaplanan

yiik

sek

aynm

giicii (0

.

99999)

nedeniyIe

,

5

STR

sisteminin birlikte bu yorenin adli

ama~lI

DNA

anaIizlerinde

kullamlabilecek

ge~erli

genetik i§aretler oldugu dii§iiniiImektedir.

Anahtar kelimeler

:

STR, HumTHOl, HumvWA, HumFI3B, HumCSFJPO,

H

umD21

S

11,

DNA

analizi,

PopuJasyon

~alI§masl.

Summary

Allele Frequencies Distribution of

5

STR Systems

in

t;ukurova

in

Region

In

this

study

,

allele frequencies

for the

STR

lo

c

i

H

um

TH

Ol,

HumvWA

,

H

umFI3B,

H

umCSFIPO and

H

umD21S11 were detern]ined in 110

Turkish

population s

amples

living

in

9ukurova

region.

It was

evaluated that

these

STR

loci

could

be

valuable

for forensic

iden-tification and paternity and

the results

were compared

with

other population

studies.

Blood samples collected

from unrelated healthy donors were dried on

a

W

ha

tman

n

fil-ter paper.

Prepared bloodstains

were extracted using

C

helex

extraction method.

Extrac

ted

DNA

samples were

amplified

by thermal cycler

and

amplified

products were

separated on

6%native

PAG,

visual

ised

b

y s

ilver

staining.

Seven alleles (6-10) were

i

d

ent

ified for

THO

l

,

eight alleles (13

-2

0) for

vWA,

fiv

e

alle-les (6

-

10) for

F

I

3B, s

ix

alleles (9-14)

for

CSFIPO and

twelve

alleles (26-34.2)

for

D2

I

S

l1

.

N

o deviation from

H

ardy

-We

inberg

Equilibrium

was found.

Because

of

the hi

g

h di

s

c

r

imi

-nation power (0

.

99999), it is th

o

ught that these five

STR

syst

e

ms have valuable

ge

n

e

tic

markers for forensic

DNA

analysis in this region.

Key words:

STR,

H

umT

HO

I,

H

umv

WA

,

H

um

F

1

3B, Hu

mC

SFIPO, HumD2

I

S

l

1,

DNA

analysis

, Pop

ulation

studies.

Giri§veAm~

Sh

ort

T

andem

Repea

t (S

TR

)

lo

kuslan

insan

genomunda

2

-7

baz

~ifti

uzunlugundak

i

Adli TIp Derg., Cilt 15, SaY12, Yl12001, Sayfa 1 -5

ADLİ TIP DERGİSİ

Journal of Forensic Medicine

Adli Tıp Dergisi 2001; 15(2): 1-5

tekrar

baz dizilim1eri

is;eren

liniteleri

di

r. i

n

san

genomu boyunca her

6-10

kb

arahkla ve

yakla§lk

4x108

STR

l

o

kusunun

bulundug

u

belirti

l

mektedir (1-5).

STR

lokuslan

genetik haritamn ortaya S;lkan

l

masl, antropolojik s;ah§malar

ve adli

amas,:h

k

im

liklendinne

ve patemite

tayininde

kullam

l

maktadlr

. PCR

i

l

e etkin amplifIkasyon

ozelligi az

mikta

rda

ve/veya pars;al

anm

l§

DNA

i<;;eren biyolojik materyaller

in

adli

analiz-l

erinde kolaybk

s

a

gla

maktadl

r

(1

-5).

Genetik

i

§are

t

lerin kimliklend

i

nne

ve akrabahk ili§ki

l

erinin tayininde

kulla

m

mmdan

once, herhangi bi

r

ge

n

et

i

k i§aretin

i

l

gil

i populasyonda allell

f

rek

ans dagIllmmm

hesa

p

lan-m

a

sl

ve ilgili

populasyo

n

is;in

adli

e

tkinli

k

degerlerinin

b

elirlenmesi

gerekmekte

d

ir

.

Bu

amas;la s;ah§mam

Izda THO

l

,

vWA,

F13B

,

CSFIPO

ve

D21S11

STR

l

o

k

uslanndan

o

l

u§an

5

s

istem

in

<;

u

kuro

v

a yoresinde allellfrekans dagIllm

o

ranlan

ara§

tl

n

l

ml§ ve

adli etki

n

lik

degerlerinin

i

statistiksel

olarak kimliklendinnede

a

nlamh

olup o

l

madlgl tartl§Ilml§tlr.

Aynca elde edi

l

en

s;a

h§

ma s

o

nus;lan

s;e§itl

i tilke

l

er

i

s;

in b

ildiri

len diger pop

u

la

syon s;ah§ma

s

on

us;lan ile kar§l1a§tlnlml§trr (6-9)

.

D

egerlendinneye alman lokus

l

ann b

u

l

u

nd

u

gu kromo

z

om, tet

r

amerik s;ekirdek

d

izisi ve

ilgili

gen bankas

l

numaraSl

Ta

b

lo l'de goste

r

i

l

mektedir (3,4,10

-13)

.

Tablo 1.

(:alI§

11

an

PoiimoIflk

STR Lokus/an.

LoJrus

Te1::rar

DiD

HumTHOI

AATG

HumvWA

AGAT

HumF

13B

AAAT

HumCSF

I

PO

AGAT

HumD21S11

AGAT

Materyal ve

Metod

Kromozom

llp15.5

1

2p

1

2-p

ter

lq31-q32

.1

5q33.3

q34

2

1

ql1-21q21

Genbank1tasl

numarasl

D00

269

M25858

M

64554

X

1

472

0

M84567

<;ukurova

yoresinde ya§ayan ve ara

l

annda akrabahk

i

li§k

isi

b

ul

unmayan

110

gonlillti

ve

riciden

alman venoz

kan

omeklerinden whatmann f

i

l

t

re

kagidl lizerine

l

eke

1e

r

ol

u§turul-d

u

o

O

l

u§turulan

lek

e

lerd

en

Chelex

ekstraksiyon metodu ile

DNA

e

k

strakte edildi

(1

4). PCR

reaks

iyo

n

k

an§lm

l

ve a

m

pl

i

fikasyon §artlan

i

malatS;

l

finnan

m

(

Pro

mega

Corp

o

rat

ion)

onei-i

leri dikkate almarak

gers;ekle§tir

ildi.

PCR

liriinleri native hor

izont

al

poliakrilamid jel

elek-troforezi

i

le a

yn§tlnidl ve gtimli§ boyama yontemi

ile g

orlinli

r1e§t

iril

d

i

(15-17).

A

lle

l

d

i

zaym

ve adlandmna,

U

lus

la

raras

l Adli Hemogenet

i

k Demegin

i

n oneri

l

eri

dikkate

a

l

marak yapIldl (18)

.

Ortala

ma Dl§lama

~ansl

is;in Wiegand

(19

)

ve arkada§lannm;

aylrt

etme

glicli

,

het

erozigotluk

oram ve standart sapma

is;i

n

Fi

sher (20

)

ve

arka

da

§lan

ile

Edwards

(2

1)

ve

arka

d

a§

l

annm kullandIg

l

istatistiksel yontemlerden yararlamldl.

Elde

e

dilen

verilerin

Hardy-Weinberg E§itligine uyumu bas

it x

2

_{test veya}

_Rand

₍₂₂₎

_ve

arkada§

l

anmn kullandlgl binning

yakla§lm ve

HWE

software

pake

t

pr

og

ramlanyla

hesap-landl

(

23

).

Sonu~lar

ve 18rl1§ma

<;ukurova yoresinde

ya§ayan ve

ar

al

armda akrabah

k

ili§kisi

tulunmayan

110 ki§iden

a

l

man kan omeginde

5 tetramerik

STR lokusunun

fenotipleri

ile adli etkinlik degerleri

ara§tmldl.

<;ah§ma

yapllan

5 STR lokusunun

allel

frekanslan ile homozigot

ve heterozigot

ki§i

saylSl

Tablo

2'de

gosterilmektedir.<;ah§mada TH01 is,:in

7 allel (6-11), vWA

is;in 8 allel

<;:ukurova Yiiresinde 5 STR Sistemi (HUMTJ-iOI, HUMvWA, HUMFI3B, HUMCSFIPO, HUMD2IS11),nin Allel Frekans Daglhml

(13-20),

F13B is;in 5 aBel

(6-10),

CSFIPI is;

in

6 aBel

(9-14)

ve

D21S11

i

s;in 12

aBel

(26-34

.

2) saptanml§tlr.

Tabla

2. <;ukurova

yoresinde

5

STR

sisteminin allel frekans

dagillmi

(AF:Allel frekansl).

HumF13B

HumCSF1PO

HumTIlOl

HumvWA

HumD21S11

Homozigot

34

Homozigot

28

Homozigot

26

Homozigot

27

Homozigot

19

Heterozigot

76

Heterozigot

82

Heterozigot 84

Heterozigot

83

Heterozigot

91

Allel AF

n

A11el AF

n

Allel AF

n

Allel AF

n

Allel AF

n

6

0.091

20

9

0.036

8

6

0.286

63

13

0.009

2

26

0.005

1

7

0.005

1

10 0.336

74

7

0.159

35

14

0

.

082

18

27

0.014

3

8

0.327

72

11

0.227

50

8

0.095

21

15

0.068

15

28

0.182

40

9

0.205

45

12

0.336

74

9

0.268

59

16

0.232

51

29

0.214

47

10

0.373

82

13

0.050 11

9.3

0.164

36

17

0.355

78

30

0.214

47

Total 1.000

110

14

0.014

3

10

0.023

5

18

0.182

40

30.2

0.023

5

Total 1.000

110 11

0.005

1

19

0.064

14

31

0.068

15

Totall.000

110

20

0.009

20

31.2

0.136

30

Total 1.000

110

32

0.009

2

32.2

0.100

22

33.2

0.027

6

34.2

0.009

2

Total

1

.000

110

Eide edilen verilerin Hardy-Weinberg E§itligi ile uyumlu oidugu bulundu p(O.05).

Saptanan

THOl, vWA

aBel frekans

l

an Tiirkiye'de

yapllan diger s;ah§ma

sonus;lan

i

l

e

kar§Ila§tln

ld

lgm

d

a

a

B

e

l

frekans

dagIllm

l

annda

kiiS;

iik

farkh

hk

lar

oldugu gozlenmektedir.

A

vrupa populasyon s;ah§ma so

n

us;

l

an

il

e yapIlan k

ar

§

l

l

a

§

tlrmala

rda THO

1

, vW A,

F13B ve

CSFIPO aBel frekans

d

agIll

m

lan arasmdaki

f

a

r

k

hhklann,

D21S

1

1

lo

k

usu ile k

ar

§lla§tml

d

l-gmda daha dii§iik de

r

ecede oldugu gozlend

i

. K

ar§Ila

§tJnna Uzak Dogu

iilkeleri

i

le

yapIldl-gmda goz

l

enen farkhhgm s;ok daha faz

l

a oldugc goz

l

endi

(3,6

-8,24-32).

T

ab

lo 3'te 5

STR

sistemi

n

i

n

gozlenen

ve

b

ek

l

enen heterozigotluk

o

ranlan,

ortalama

dl§lama §ansl ve aYlrt

etme giic

iin

de

n

olu§an adli etki

n

l

ik

degerleri

goriilmektedir.

Degerlendirmeye alman lokuslardan en kiiS;ii

k

aYlrt e

t

me giiciine sahip lokus %85'le F13B

,

en yiiksek ay

l

rt etme giiciine sahip lokus %95'le D2

1

S

1

1'dir. Ancak ses;ik

n

lo

k

uslann

stan-dar

d

ize e

d

i

l

ebilme avantajl

dikkate ahndlgm

d

a;

F13B

lokusunun aylrt etme

giicii

dii§iik

olmasma ragmen standard

i

zasyonu

k

o

l

ay o

l

dugundan rutin uygulamada terc

i

h sebebi o

l

arak

kabul edilebi

l

ir.

Tabla

3.

<;ukurova yoresinde

5

STR sistemi iqin belirlenen

adli

etkinlik degerleri (H-Obs:

gozlenen

heterozigotluk;

H-Exp:

beklenen heterozigotluk; MEC:

ortalama

dl§lama §ansl;

DP:

aylrt

etme

gilcil).

Lokus

H-Obs

H-Exp:tSD

MEC

DP

HumTHOI

0.7545

0.7877±0.0201

0.57691

0.91752

HumvWA

0.7182

0.7755±0.0294

0.56822

0.92149

HumF13B

0.6909

0.7071±0.0248

0.44508

0.85967

HumCSF1PO

0.7455

0.7213±0.0249

0.46852

0.86281

HumD21S11

0.8000

0.8446±

0.0173

0.68233

0.95107

Kombine

0.99999

Yarg

lt

ay II. Huk

uk

Dairesi

n

in

1

5.11.1993

t

arih ve 9405

ka

r

ar no'lu iIammda babahgm

%99.73'ten daha az

iht

i

maBe kabuliin

iin

m

iimkiin olamayacagi be

lirti

lmektedi

r

(33).

Ta

b

lo

3'te aynca bu 5 STR lokusunun

k

om

b

inasyonu ile

0.99999'luk

bir aylrt etme

giiciine

olarak,<;ukurova yoresinde

adli

ama'rh kimliklendinne ve

akrabahk ili§

ki

lerinin tayininde

kullan~labilecek

etkin i§aretler oldugu kabul ed

i

lebilir.

Kimliklendirme ve patemite tay

i

ni ic;:in kullalll

l

an polimo

rf

ik sistem ne kadar art

m

hrs

a

,

o oranda dogruya en yakm istatistiksel

ve

r

i elde edilebilmektedir. Ulkemiz

ko§u

ll

armda

genellikle

ana

kan grup ve subgruplarl ile HLA antijenlerinin analizi sonucu

elde e

d

i

l

en

veriler degerlendirilmektedir. Anabilim Dahmlzda ara§tIrmasl yapIlan patemite olgulannda,

b

u c;:ah§mada belirtilen 5 STR sisteminin rutin uyg

u

lamaya konulm

a

SI

ile sonu'r ahna

m

aya

n

olgu

analizlerinin

c;:ogunda

b

i

z

i

anlamh sonuca

ul

a§tIrabilecek bi

l

gi

l

erin yamnda; <;e§itl

i

biyolojik leke ve kahntIlann da tammlanmasl miirnldin o

l

dug

u

ndan,

aynca kimli

kl

en

di

rme

'rah§malannda ku

ll

alll

l

abilecektir.

Geli§mi§

iilkelerde

geleneksel sistem1erle (hiicre

antijenle

r

i,

p

o

limor

f

i

k

e

n

zim

ve

pro-tein1er)

b

irlik

t

e kullallllan DNA ana

l

izlerinde veri

k

ar§l1a§tIrmasl y

a

pllama

d

lgl ge

r

ekc;:esi i

l

e

kulla

lll

lan sis

t

emler ve sec;:

il

en yontemler

iizerinde standa

r

dizasyon belirlenmeye

c;:ah§Ilmak-tadrr. Ya§anml§

o

l

an deney

i

mler

d

ikkate

a

l

marak, tilkemizde far

kll

bir

i

m ve

bolge

l

erde

popu

l

asyon c;:a

h

§masl yaplldlktan soma rutinde aylll polimorfi

k

sistem ve yontemlerin

sec;:i-m

i el

d

e edilen verilerin daha verimli kullallllabilmesine olanak

t

alllyacaktrr. A

n

cak aylll

sis-tern ve

yontem kullammmda

dahi

f

arkh laborat

u

var sonw;larmm

ortaya

'rlkabi

l

ecegi

goz

onii

n

de bulundurulmahdlr (34,35)

Kaynaklar

1. Urquhart, A., Kimpton. c.P., Downes, TJ .• Gill, P. (1994) Variation in short tandem repeat sequences- a survey of twelve microsatellite loci for use as forensic identification markers, Int. J. Leg. Med. 107, 13-20.

2. Kimpton, c.P .• Fisher. D, Watson, S., Adams. M., Urquhart. A., Lygo, J., Gill. P. (1994) Evaluation of an auto mated DNA profiling system employing multiplex amplification offour tetrameric STR loci. Int. J. Leg. Med. 106, 302-311.

3. Hammond, H.A., Jin, L., Zhong, Y .• Caskey, C.T., Chakraborty, R. (1994) Evaluation of 13 short tandem repeat loci for use in personal identification applications, Am. J. Hum. Genet. 55, 175-189.

4. Hochmeister, M.N., Budowle, B., Schumm, J.W., Sprecher, CJ., Borer, U.V., Dimhofer, R. (1995) Swiss popula tion data and forensic efficiency vaiues on 3 tetrameric short tandem repeat loci - HumTHO 1. TPOX and CSFIPO-derived using a STR mUltiplex system. Int. J. Leg. Med. 107,246-249.

5. Huang, N.E., Schumm, J., Budowle, B. (1995) Chinese population data on three tetrameric short tandem repeat loci -HumTHOI, TPOX and CSFIPO-derived using multiplex PCR and manuel typing. Forensic Sci. Int. 71, 131-136.

6. Alper, B.. Meyer, E., Schurenkamp, M., Brinkmann, B. (1995) HumFES/FPS and HumFI3B: Turkish and German population data, Int. J. Leg. Med. 108.93-95.

7. Alper. B., Wiegand. P., Brinkmann, B. (1995) Frequency profiles of 3 STRs in a Turkish population, Int. J. Leg. Med. 108. 110-112.

8. Iwasa, M .• Wiegand, P., Rand. S., Schurenkamp. M., Atasoy, S., Brinkmann, B. (1997) Genetic variation at five STR loci in subpopulations living in Turkey, Int. J. Leg. Med. 110,170-172.

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Ktsal1malar:

STR: Short Tandem Repeat PCR: Polymerase Chain Reaction PAG: Polyacrylamide Gel DNA: Deoxyribonucleic acid HWE: Hardy-Weinberg Equilibrium

"'Bu ~alI§ma 22-25 Eyliil 1999 tarihinde Almanya-Mainz'da yapllan IV. International Symposium Advances

in Legal Medine'da ve 10-13 MaYls 2000 tarihinde istanbul'da yapllan IV. Adli Bilimler Kongresinde POSTER

olarak sunulmu§tur.

Ueti§im adresi: Do~.Dr. Behnan ALPER <;:ukurova Oniversitesi TIp Fakiiltesi Adli TIp Anabilim Dah

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