FORENSIC GENETICS

FORENSIC • PERSONAL IDENTIFICATION

GENETICS
• KINSHIP ANALYSIS

FORENSIC GENETICS FORENSIC GENETICS

• Establishing human corpse identity
• creating population data frequency
• Crime cases – matching suspect with evidence used for evaluation of proof value of case report

• Paternity testing, even after death • establishing databases of convicted felons

used for linking unsolved crime cases with serial
• Missing persons investigations offenders

• establishing databases of missing persons

• Putting together tissues coming from particular
used for identification of unknown human remains
persons in mass disasters

Sources of biological evidence DNA in the cell

• Blood
• Semen
• Saliva
• Urine
• Hair
• Teeth
• Bone
• Tissue

Somatic cell
Hair root 3-6 pg DNA DNA profile
Sperm cell (0.000000000003 g DNA)

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 Cells without nucleus DNA in the Cell
chromosome

cell nucleus

double stranded
DNA molecule
A T G C

Individual
Mature red blood cells, hair shaft nucleotides

HUMAN GENOME = 3 bilion base pairs

Principles of Mendelian inheritance

The law of segregation

Each individual has two alleles lying opposite
to each other in the pair of homological chromosomes,
one inherit from the mother and second inherit
from the father

The law of independent assortment

The pair of alleles coding one marker is independent
of the pair of alleles coding another marker

DNA POLIMORPHISM Variable number of tandem repeat (VNTR)

Lenght polymorphism 10-100 nucleotides = minisatellites

VNTR - Variable number of tandem repeat
AATGGCTCTTATGACGTATCATGACTAG

Sequence polymorphism

SNP - Single nucleotide polymorphism

The number of repeats can differ between individuals

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 Variable number of tandem repeat (VNTR) Variable number of tandem repeat (STR)

AATG AATG AATG

2-6 nucleotides = microsatellites = STR (short tandem repeat)

9 repeats
AATG AATG AATG

5 repeats

locus allele 5,9 genotype 5/9

Heterozygote = the genotype at the locus has two different alleles
The number of repeats can differ between individuals

Variable number of tandem repeat (STR)

AATG AATG AATG

7 repeats

7 repeats

locus alleles 7,7 genotype 7/7

Homozygote = the genotype at locus has two copies of the same allele

DNA detection in capillary sequencer

Schema of a gel electrophoresis system

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 MPS – massive parallel sequencing VNTR analysis in paternity testing

1 2 3 M 1 2 3 M

1-allaged father 2- child 3-mother M –height marker

STR analysis in paternity testing Paternity exclusion principles

Arising from Mendelian inheritance rules

New feature is present in the child,

while it is absent in the alleged father
and the mother

and or

Without Paternity The child does not inherit any feature

exclusion exclusion from the alleged father

PCR – Polymerase chain reaction Multiplex PCR

Enables to produce millions of copies Multiple STR regions are examined simultaneously
of a specific DNA sequence in approximately two hours

Multiplex PCR reaction mix contains

multiple primer pairs

even more than 20 primer pairs

Particular primer pairs are labeled

with different fluorescent dyes

Classical PCR reaction mix contains template DNA, polymerase,

deoxynucleotides and a pair of primers – forward primer and reverse one

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 STR Genotyping after fluorescent detection

Advantages of Multiplex PCR

Reduces template required

Reduces labour to obtain results

Anables analysis of degraded DNA

Makes more effective analysis of mixtures

Increases the informativeness of the DNA test

Y- CHROMOSOME MARKERS Y-STR haplotyping after fluorescent detection

specific only for man

Haplotype Y - Genetic variation at multiple points along the Y chromosome

Male – specific Y- STR markers

Why the Y-Chromosome?
98% of violent crimes is caused by men

Applications

– identificaton of male or male components in mixture

– istablishing paternity for male offspring
– paternal lineages testing
– male gender identification

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GENDER IDENTIFICATION DNA POLIMORPHISM
WITH
AMELOGENIN
Lenght polymorphism
VNTR - Variable number of tandem repeat

AMG Y (112 bp)

AMG X (106 bp)
Sequence polymorphism

SNP - Single nucleotide polymorphism

Single nucleotide polymorphism (SNP)

Single nucleotide polymorphism (SNP)

Homozygote G G
Homozygote A A
Heterozygote G A

We differ from each other in 1 bp out of every 300 nucleobases

Advantage of SNP over VNTR markers Polymorphism of mtDNA

0
16365 73 HV1, HV2 – variable control
are abundant and common in the human genome
70 regions type of SNP
have low mutation rate ANDERSON SEQUENCE

due to the small molecular weight they are very useful in HV1 HV2
analysis of high degraded DNA samples Extractions of DNA
Limited & Old & Degraded
samples as in tissues such as
15 STR = 50 SNP 340
16024 - bones & teeth
0,2 ng 0.02 ng
- skeletal remains
16 569 bp - hair shafts !

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 NON RECOMBINATON MARKERS
HAPLOID MARKERS

Mitochondrial DNA Y-chromosome DNA

Uninparental inheritance
Inheritance from only one parent

PASSED DOWN
from mother to her child from father to his son

LINEAGES MARKERS

Steps involved in processing forensic DNA samples

DNA
Extraction FORENSIC DNA EVIDENCE
Evaluation of DNA INTERPRETATION
quantity and quality

PCR amplifiation Kinship Analysis

multiple STR/SNP
markers paternity testing

Separation and
detection of
DNA alleles

PE for first degree expertise as well as DNA expertise

PE - POWER OF EXCLUSION
DNA EXPERTISE PE=99,9999%
100%

It is the percentage of the unjustly sued man who will be 95%

excluded as fathers in the course of investigation
90% STR-Multiplex
PE determines usefulness for paternity testing 15 loci
I º EXPERTISE PE=84%
85%
ABO PGM
Kell ACP
The more polymorphic the investigated markers are, the easier it is 80% MN GLO
to exclude paternity of the man using these markers Rh ESD
GM HP

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 AN OPINION IN PATERNITY TESTING AN OPINION IN PATERNITY TESTING

DNA EXPERTISE
Iº EXPERTISE

EXCLUSION CONFIRMATION
EXCLUSION LACK OF EXCLUSION ? with 4 ( four )
INDEPENDENT MARKERS
probability of patrernity = 99.9999 %
RECONSTRUCTION OF GENOTYPE OF DEAD PERSON
International Society of Forensic Genetic
Polish Society of Forensic Medicine and Cryminology

RECONSTRUCTION OF GENOTYPE OF DEAD PERSON

Paternity testing evidence

FORENSIC DNA EVIDENCE

Paternity Probability of INTERPRETATION
Index (PI) Paternity

10 90 %
100 99 %
1000 99.9 % personal identification
10 000 99.99 %
100 000 99.999 %
1 000 000 99.9999 %

Requirements for issuing a report with

a positive weight for paternity

PD IN PERSONAL IDENTIFICATION
PD - POWER OF DISCRIMINATION

ABO D16S539
Rh D3S1358
Probability that two unrelated individuals will have KELL D2S1338
different sets of genetic alleles MN D8S1179
GM D18S51
ESD D12S11
PD determines usefulness for personal identification ACP D19S433
PGM vWA
GLO TH01
The more polymorphic markers are examined, the greater the HP FGA
chance that two unrelated individuals will not have an identical set
of these markers
PD ≈ 95 % PD = 99.9999999 %

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 GENETIC IDENTIFICATION CALCULATION OF PROFILE FREQUENCY

Comparative analysis of If allele in locus A has frequency a

received DNA profile and allele in locus B has frequency b

taking into account

assumption of HWE

No match Match
Exclusion Inclusion
AA = a2
Genotypes frequency BB = b2
If match occurs, Generation of case report
calculation of DNA profile with random match AB = 2ab
frequency probability

EVIDENCE VALUE OF PERSONAL IDENTIFICATION

1
= 1 PROFILE in .... persons
PROFILE FREQUENCY

Product of multiplication genotypes frequency

= Profile frequency PF
RANDOM MATCH PROBABILITY

The chance that a randomly selected individual from

PF = GF1 marker x GF2 marker x GF3 marker x .......
a population will have an identical DNA genotype
such as another individual in the population

RANDOM MATCH PROBABILITY USING 13 LOCI

DNA MOLECULAR
POLYMORPHISM DACTYLOSCOPY