DNA Profiling in Human Identification

Bhinu Shova Tuladhar

MSc.(Chem), MSc.(Forensic Science)
 DNA ?
DNA (Deoxyribonucleic acid) is the genetic material of all
living organisms.

It can make copies of itself and carry on the same information

to build the machinery of life.

DNA consists • 3 components:

• Sugar
• Phosphate
• Base

• 4 bases:
• Adenine ( A)
• Cytosine (C)
• Thymine (T)
• Guanine (G)
Structure of DNA
DNA is a double-helix and has two strands running in opposite
directions. Each chain is a polymer of subunits called
nucleotides .
 Genetic information in DNA is encoded as Adenine(A),
Thymine (T), Cytosine (C) and Guanine (G) 4 different
bases within the DNA structure.

Each human beings possesses specific and unique

DNA sequences.
5’….AATTGCCCCGGGAA……TAAATCTGGGCCCA…3’

3’….TTAACGGGGCCCTT…….ATTTAGACCCGGGT….5’

DNA has Coding and non coding (Junk DNA) genes.

Non-coding region is highly variable and found variable
between individual to individual.
Human genome
Nuclear genome (3X10 9 bp)
Mitochondrial genome (16569 bp)
Location of Nuclear DNA
Nuclear DNA is tightly packed
in 23 pairs of chromosomes.
Out of which one pair is sex
chromosome and 22 pairs are
homologous chromosome which
are known as autosomes.
Female have homologous X
chromosome and male have
one X and another Y
chromosome.
Importance of DNA evidence in crime investigation
 DNA is a powerful investigative tool, with the exception of
identical twins, no two people have the same DNA. In other
words, the sequence or order of the DNA building blocks is
different in particular regions of the cell, making each
person's DNA unique.
 DNA evidence collected from a crime scene can link a
suspect to a crime or eliminate one from suspicion in the
same way that fingerprints are used.
 If a biological sample from the victim, such as a bloodstain
left at a crime scene, the DNA taken from that evidence
can be compared with DNA from the victim's biological
relatives to determine if the bloodstain belongs to the
victim.
 When a DNA profile developed from evidence at one
crime scene is compared with a DNA profile developed
from evidence found at another crime scene, they can be
linked to each other or to the same perpetrator, whether
the crime was committed locally or in another state
 DNA evidence in the form of saliva, blood, skin tissue,
hair, and semen recovered from crime scenes can be
crucial to the investigation of sexual assaults and other
violent crimes. This evidence is helpful in proving that
there was physical contact between an assailant and a
victim.
 When investigating sexual assault or rape cases, it may
be necessary to obtain an elimination sample, such as a
blood or saliva sample, from the victim's relatives or
consensual sex partner.
 Demand DNA test ?
 To identify the potential suspects by matching
crime scene evidence with suspects and link
him or her to a particular crime (Murder, Rape, Assault,
Robbery, Kidnapping, etc).
 To identify crime and catastrophe victims.
 To establish biological relationship of persons.
 To exonerate persons wrongly accused of
crimes.
 To identify endangered and protected species of
Wildlife poaching.
 Mass disaster investigation cases.
Issues that can be resolved through DNA test

Mass disasters
Identification of
missing persons
Wildlife poaching and illegal trade of wildlife
 Sources of Biological Evidence

Blood Saliva Semen Tissue

Hair Bone
Teeth Urine

All above sources contain DNA and found same i.e.,

DNA in blood is same as the DNA in skin cells, saliva,
hair etc.
DNA profiling
 Introduced by Sir Alec Jeffery‘in 1985.
 Reliable and confirmative human
identification system.
 DNA profiling of each person is unique except
identical twins.
 Protect the interests of innocent persons in
criminal cases.
 Currently used in the investigation of crimes
and disputed paternity.
 Aim of DNA Profiling
Produce a profile that can: Crime Scene

link a person to a crime

link biological relatives.

How ?
DNA polymorphisms allow the possibility to
generate a unique profile. bb
 Currently used Human Identification systems in forensic
cases

 PCR based STR profiling of chromosomal DNA in the

nucleus is the most powerful and robust contemporary
technique for human identification. The accuracy of the
PCR based STRs DNA profiling technique is 99.9999 %.
 mtDNA typing is resorted to, When STR DNA typing
could not be done due to degradation of biological
specimen. It provides an advantage over nuclear DNA
for solving forensic cases involving missing persons.
 Y-STR typing is used in rape and sexual assaults
cases involving single or multiple assailant. It is a
strong evidence for paternity within a male line
spanning many generations.
 Short tandem repeats
STRs (STRs
are )
locations (loci) on the chromosomewhich
contain short sequence fragments (2-7 bp) that repeat
themselves within the DNA molecule.
Usually tetra nucleotide repeats AGAT or GATA or AATG
are used in forensic identity testing.
AGA
T

Allele 1
6 repeats
Allele 2
8 repeats
___AGAT AGAT AGAT AGAT AGAT AGAT____
___AGAT AGAT AGAT AGAT AGAT AGAT AGAT AGAT____
the repeat region is variable between individual to individual
Which made them important to use them in human identity testing
 16 STR Loci with
Chromosomal Positions
TPOX

D3S1358

TH01
D8S1179
D5S818 VWA
FGA D7S820
D2S1338 CSF1PO

AMEL
D19S433
D13S317
D16S539 D18S51 D21S11 AMEL
Accuracy of matching DNA profile in people depends on the
number of loci checked for STR alles. The greater the number of
STRs produce a higher the accuracy of the identification.

No. of STRs Frequency of occurrence in range

3 1 in 5000 (thousand)
6 1 in 200,0000 (million)
9 1 in 1000,000,000 (billion)
13 1 in 1000,000,000,000 (trillion)

SGM plus ( 10 STRs) Used in UK.

CODIS (Combined DNA Index System) (13 STRs) Used in
FBI , USA
AmpFISTR identifier (15-STR s) currently used in most of the
countries.
Mendelian Inheritance
Example
Mother Father

4,8 5,12

Child
4,5 4,12 5,8 8,12
Preserving & Collecting Forensic Evidence in Sexual Assault case
DNA evidence is an integral part of a law enforcement investigation that can
build a strong case to show that a sexual assault occurred and to show that
the defendant is the source of biological material left on the victim’s body.
 the most important thing is for the victim to get to a safe place.
 it is vital for the victim to receive medical attention, and strongly
recommended for the victim to receive a forensic examination
Victims should make every effort to save anything that might contain
the perpetrator’s DNA, therefore a victim should not
Bathe or shower
Use the restroom
Change clothes
Comb hair
Clean up the crime scene
Move anything the offender may have touched
Even if the victim has not yet decided to report the crime, receiving a forensic
medical exam and keeping the evidence safe from damage will improve the
chances that the police can access and test the stored evidence at a later
 DNA Capabilities and Experiences
o NAFOL is a pioneer
o Laboratory which has DNA analysis facilities
available in Nepal.
o Since 2006 the laboratory has been providing
DNA analytical services to establish biological
relationship in disputed paternity, immigration,
citizenship and other cases.
o The laboratory uses highly confirmative, reliable
and widely accepted PCR based STR DNA
profiling Technique.
 DNA Capabilities and Experiences
Cont…

 Started Y-STRs DNA profiling for father line

relations ( Grand father-father- Uncle- Son
etc.)

 STRs DNA profiling of human remains( bones,

teeth etc.) technique developed.
 mtDNA sequencing in wild life forensic and
missing identification.
 Steps in DNA sample Processing
Samples obtained from crime scene or
paternity investigation

DNA DNA PCR amplification

extraction quantitation multiple STR

Separation and detection of PCR

products (STR Alleles)

Sample genotype determination

Comparison of sample genotype to other sample results
 Puncture of 1.2 mm FTA blood
Collection of blood in FTA card stain

Transferring 1.2 mm FTA blood stain into

PCR tube and purification Purified DNA
Extraction of DNA from bone sample
 Amplification of DNA samples in PCR

PCR
(Polymerase Chain Reaction)
Getting DNA
profile using
DNA Analyzer
ABI 310

Electropherogram
Allele analysis in the DNA profile of the samples
S.N Locus Mother Child Alleged Remark : Unmatched child
Father alleles w.r.t. Alleged Father
1 D8S1179 10. 10 10, 10 14, 16 10
2 D21S11 28, 32.2 30, 32.2 29, 30 -
3 D7S820 12, 13 11, 12 11, 13 -
4 CSF1PO 8, 13 8, 12 11, 12 -
5 D3S1358 16, 18 16, 17 17, 18 -
6 THO1 6, 9 9, 9 6, 7 9
7 D13S317 12, 13 10, 13 8, 13 10
8 D16S539 9, 9 9, 13 12, 12 13
9 D2S1338 21, 24 21, 24 22, 23 24
10 D19S433 13.2, 15.2 14, 15.2 13, 15 14
11 vWA 15, 16 16, 16 17, 17 16
12 TPOX 8, 10 8, 10 8, 10 -
13 D18 S 51 14 ,14 14, 20 15, 15 20
14 D5 S 818 10, 12 11, 12 11, 13 -
15 FGA 20, 21 21, 24 22, 25 24
16 AMEL XX XY XY -
Three Possible Outcomes
 Inclusion (Match) – Peaks between the
compared STR profiles have the same
genotypes .

 Exclusion (Non-match) – The genotype

comparison shows profile differences .

 Inconclusive – The data does not support a

conclusion (insufficient information exists to
support any conclusion)
Match? Issue ?

 How often would a random unrelated person be

found with that particular DNA profile?
 Probability ? (the chance of a particular event
occurring)

 What is the statistical calculation for the

probability of there being a random match on
these loci?
DNA Database

Population databases of major racial and ethnic groups are

used to determine estimates of the rarity of DNA profiles.
The Reality
Once an individual's 13-locus STR profile is identified, it is
statistically improbable that anyone else in the world will have
the same profile, unless that person has an identical twin.

All 13 CODIS sites would mean that the chances of matching

a random DNA sample are about one in ten trillion.

It means probability of two different people matching at all 13

CODIS sites is virtually zero
DNA Data Bank
Indexing of DNA profiles of forensic cases in
National DNA Database.
Principle of a DNA Database
1)DNA obtained from the genetic material of a
person or from biological evidence that is left at a
crime scene is available on a database for
investigation purposes.
2) DNA from one source will give the same profile
whether it comes from a crime or a reference
sample.
 Challenges

1.Issues involving contamination.

(Mixed DNA profiles may lead inconclusive results and
contamination does not change a person’s DNA)

2.Degradation of DNA
(Difficult to get DNA extract and may not get DNA profile.
Degraded sample would not produce a false result rather no
results or partial results.
Degradation does not change person’s DNA.)
Successful Crime investigation ?

Admissibility of DNA evidence in a court.

what are necessary requirements?
Forensic sample handling and collection should
meet the legal and scientific requirements.
1) Legal : well documenting
2) Scientific: collecting, packing and preserving
3) Chain of Custody: securing evidence by sample
tracking.
 If DNA evidence is not properly documented, its origin
can be questioned.
 If it is not properly collected, biological activity can be
lost.
 If it is not properly packed, contamination can occur
 If it is not properly preserved, decomposition and
deterioration can occur.
Issues raised ? Laboratory able to satisfy ?

Chain of custody  Requested letter from court or

any authorized organization.
Sample handling ?
 Sealing of sample with unique
authentic impression.
 Record of handling of samples.
Storage before analysis Analysis
Final state of sample
(Store or return)
Issues raised ? Laboratory able to satisfy ?

Contamination Source of contamination.

a) sample collection.
Contamination (can not recovered)
free How ? b) Laboratory work.
(recovered by retest)
• All personal precautions
( wearing gloves, mask,
protective clothes etc.)

• Careful handling of the

samples
 Issues raised ? Laboratory able to satisfy ?

 The test methods used are

Reliability of tests confirmative, reliable and
validated internationally
Assurance in validation
and reliability of results.
 widely world wise accepted
techniques.
How ?
(The accuracy of the PCR
based STRs DNA profiling
technique is 99.9999 %.)
 Participating in International
DNA Quality Control
proficiency testing programs.
 The alleles investigated at 15 STR loci from the crime scene sample
and the two suspects (theft case), and from the missing man, son of the
missing Man, crime scene samples and the suspect (murder case)

Allele designations Theft case Murder case

Killed
Locus Crime Suspect Suspect missing Son of Crime scene Crime scene Suspect
scene sample 1 2 man missing sample 1 Sample 2 man

D8S1179 13,14 14,15 13,14 13,15 13,14 13,15 10,14 10,14

D21S11 29,29 29,32.2 29,29 28,31 28,31.2 28,31 29,30 29,30
D7S820 8,10 10,11 8,10 8,11 11,12 8,11 8,10 8,10
CSF1PO 11,13 10,11 11,13 10,11 10,11 10,11 10,10 10,10
D3S1358 16,17 15,15 16,17 14,16 16,17 14,16 17,18 17,18
TH01 6,6 9,9.3 6,6 7,7 7,9 7,7 7,9 7,9
D13S317 8,9 12,12 8,9 11,11 11,12 11,11 11,14 11,14
D16S539 13,13 9,12 13,13 11,11 11,11 11,11 9,11 9,11
D2S1338 19,22 17,23 19,22 24,25 24,25 24,25 20,23 20,23
D19S433 13,15 14,14 13,15 13,14 14,15 13,14 14,16 14,16
vWA 16,17 17,19 16,17 14,16 16,16 14,16 16,17 16,17
TPOX 8,8 8,8 8,8 8,11 8,10 8,11 10,11 10,11
D18S51 13,16 13,15 13,16 13,16 16,18 13,16 17,18 17,18
D5S818 8,13 10,13 8,13 11,12 11,11 11,12 12,13 12,13
FGA 22,27 22,23 22,27 21,22 21,23 21,22 24,26 24,26
Amelogen XY XY XY XY XY XY XY XY
DNA Sample collection in Missing
Identification