Paternity Testing

(Whos Your Daddy?)

What is DNA?

DNA (Deoxyribonucleic acid) resides in the nucleus of eukaryotic

cells and contain all necessary information for the maintenance of
overall body function. It has the ability to transfer information to
successive generations. It also has the capability to replicate itself.
Thus, this explains why it is a superior sample for relationship or
forensic disputes.
How do I inherit traits from my parents?

DNA is established at conception, and does not change throughout your life.
You receive one-half of your DNA from your mother and one-half from your
father. DNA is what allows for the transmission of genetic material from one
generation to the next. When your fathers sperm and your mothers egg
cells combine at fertilization, they create your genetic sequence in 23 pairs
of chromosomes (46 total).
 fertilization

Fathers Sperm Mothers Egg

Childs Cell
Law of inheritance

The Mendelian inheritance law is the basis of

paternity testing.

The child inherits 23 chromosomes from the mother

and another set of 23 chromosomes from the
biological father.
 Males chromosomes

22 pairs of autosomal and a pair of sex

chromosome (X,Y)
 FEMALES CHROMOSOMES

22 pairs of autosomal and

a pair of sex
chromosomes (X,X)
A pair of homologous chromosomes,
(one from mom, one from dad)

Humans have 23 pairs of homologous chromosomes

Gene
unit of DNA information about a trait
passed from parents to offspring
specific location on a chromosome

Alleles different versions of a gene

Human Chromosomes

Fig. 8-1, p.113

What is paternity testing?

Paternity testing is DNA analysis to determine whether or not a given

man is the biological father of a given child.
Why would I want paternity testing?

There are many different reasons people want to have a paternity test.
Sometimes alleged fathers are told many years later that a previous relationship
resulted in the birth of a child, and they want to make sure they are actually the
biological father.
Some men might suspect their wives or girlfriends of infidelity and are concerned their
child may not be theirs biologically and want the peace of mind a DNA test can provide.
Children who were separated from their biological father at a very early age, whether
because of adoption or other reasons, often want a conclusive DNA test early in the
reunion process.
Children sometimes learn about infidelity after the death of a parent and are curious
about their genetic makeup and their family's medical history, so they have DNA tests
to prove their parentage.
Mothers suing for paternal recognition, support and inheritance.
How can you test DNA to determine a family relationship?

Depending on the type of relationship to be tested, the parties in

question will submit samples of their DNA collected using buccal (cheek)
swabs or blood sample.

For example, if you need a paternity test, you need to collect samples
from the child, the alleged father, and the mother (if she is available and
willing to participate). Once collected, the samples will be submitted to
the DNA testing laboratory for detailed analysis. The laboratory will then
compare specific locations on the DNA to determine whether or not
there is a biological relationship.
 Steps in doing Relationship Test
I. A pre-test conference is held
with the clients wherein they are
informed about the test
principles and processes as well
as how the results are
interpreted.

II. Clients blood or buccal swab

samples are collected by an
authorized Medical technologist
collector.
 Steps in doing Relationship Test
III. After collection, the samples
are processed for DNA extraction
and quantified to ensure that a
specified amount of DNA is
present for proper analysis.
 Steps in doing Relationship Test
IV. Extracted DNA sample
undergoes Pre-amplification using
specified genetic markers.
PCR (Polymerase Chain Reaction)
Developed in 1983 by Kary
Mullis
Awarded Nobel Prize in 1993
 Steps in doing Relationship Test
Isolate and mass produce a
particular DNA fragment
Template DNA
Polymerase enzyme that
replicates DNA, matches
complementary bases
Primers 10-30 base pairs long,
complementary to ends of
fragment
Short Tandem Repeats
Short regions of DNA that differ substantially among
people
TCAT
Each person carries a unique combination of repeats
Steps in doing Relationship Test
 Steps in doing Relationship Test
Isolate and mass produce a
particular DNA fragment
Denature
Anneal
Extend
 Steps in doing Relationship Test
V. Post amplification of amplicons
undergo denaturation where an ILS
(Internal Lane Standard) and a HI-DI
will be added to the sample.
 ILS600 Size Standard

100 200 300 400 500 600

A sizing standard is used in all samples and allelic

ladders
The known standard is used to determine the size of the
allelic ladders
and the unknown samples
 VI. Post amplified samples are
analyzed in a genetic analyzer
machine utilizing capillary
electrophoresis for data
acquisition.
 Steps in doing Relationship Test
VII. Acquired data is imported
into a Gene Mapper software.
Allelic Ladder
Allelic Ladder
PowerPlex 16
 Steps in doing Relationship Test
VIII. Analyzed data is imported into DNA View for relationship
analysis.
DNAVIEW is the universally acknowledged
authoritative world-wide standard for DNA
identification.
Leading paternity and crime laboratories
have been using it since 1988. They
appreciate not only the reliability and
sophistication of the software, but also the
authority that it represents created and
supported by Dr. Charles Brenner.
Power of Discrimination

Ability to discriminate between different individuals

The larger the number of loci used, the more powerful the ability to
discriminate
Paternity Index

Likelihood ratio: X/Y

X = chance that the AF could transmit the obligate allele
Y = chance that some other man of the same race could
have transmitted the allele

X = 1 if father is homozygous
X = 0.5 if father is heterozygous

D8 hypothetical example:
13 is obligate allele, frequency of 13 = 0.308

PI = 0.5/0.308 = 1.62
Paternity Index

1.62
12.20
4
3.60
2.28
5.68
1.40
1.82
2.89
Combined Paternity Index (CPI)

All PI multiplied by each other since the different loci are inherited
independently of each other being in different chromosomes or set
wide apart on the same chromosome

Combined Paternity Index = 27,746

Probability of Paternity

CPI
x 100
CPI + (1-prior probability)

27,746
x 100 = 99.998%
27,746 + (1-0.5)
How do I read the results of my paternity test?

A formal paternity test report contains information about the

different loci analyzed to generate the paternity result. The section
you will want to focus on is the Probability of Paternity at the bottom
of the report. This value will fall into one of two categories:
 Non-exclusion: A probability value of at least 99.99% for a trio case
(if the mother, child and alleged father were all tested) means that
the tested man is not excluded as the biological father.
EXAMPLES OF
PATERNITY CASES

NON-EXCLUSION

FATHER MOTHER CHILD

 Exclusion: A probability value of 0% means that the tested man is
excluded as the biological father.
A minimum of two loci with non-identical STR repeats between
alleged father and child
A single non-identical locus may mean a genetic mutation especially
if there is only a one repeat difference
 Genetic Mutation
Possibility exists when one STR locus eg. THO1 in the child is 8, 9
and the mother is 8,12 and the alleged father is 10, 11
There is a one repeat difference between the obligatory paternal
allele in the child (9) and the alleged father (10)
Calculations are made to determine the paternity index using
mutation rates of this locus
EXCLUSION

FATHER MOTHER CHILD

Why will results claiming probable
inclusion never equal 100%?
In paternity testing, no man will ever be 100% included as
the biological father because there is always the slight
possibility that the DNA profile of the alleged father matches
the DNA profile of the child by mere chance. The likelihood
of this happening is usually well below 0.001% (1 in
100,000), but it depends in large part on the ethnic origin of
the individuals involved. In addition, it important to note
that the certainty usually increases with the number of DNA
loci (locations) analyzed.
 What does 99.99% probability really mean?
For paternity cases, if the alleged father is not the
biological father, the result will be 100% exclusion.
However, if the alleged father cannot be excluded as the
biological father, the result will be a certain percent
inclusion. Make sure you look for a lab that routinely
provides at least 99.99% probability of inclusion. This
means that there is only a 0.01% chance that another
random individual in the same race population or more
probably, other close male relatives could have the same
paternity test results. Consider that many companies
provide 99.9999% probabilities of inclusion, meaning
there is only a 0.0001% chance that another man could
be the father.
What is a "motherless"paternity test?

A motherless paternity test can be done if the mother is unable or

unwilling to participate in the testing process. This test will provide
the same conclusive results as a standard paternity test.
 Regulation of Labs

Forensic
FBI
Standards for Combined DNA Index System (CODIS) labs
http://www.fbi.gov/hq/lab/codis/index1.htm
The Scientific Working Group for DNA Analysis Methods(SWGDAM)
publishes guidelines
Paternity
American Association of Blood Banks (AABB)
http://www.aabb.org
 Uses of DNA Testing

Main application is in Forensics to identify

Bodies of crime victims or of mass disasters
Perpetrators of crimes: murder, theft and sexual assault
Baby switching in hospital nurseries
Presence of other persons in a crime scene
Exoneration of innocent persons falsely imprisoned for crimes they did
not commit
(Project Innocence)
In Anthropology
Identification of historical figures as in mummies of Egypt and Peru
Egyptian mummies yield genetic secrets
In Anthropology
Establishing definitive identity of families as in the Romanovs of Russia
who were massacred by the Bolsheviks and buried in an unidentified
mass grave
Essentials in DNA Profiling

Crime scene investigation

DNA sample collection must document and identify the sources of these
samples as in location, type of sample: blood stains, semen, hair, fingerprints,
etc.
These must be isolated and preserved in sterile containers with appropriate
notation as above
Must have strict chain of custody to insure integrity of samples
 Essentials in DNA Profiling (2)
Testing must insure strict identification and precautions against
contamination
Testing methodology must adhere to strict standards from choice
of reagents to the testing process and instrumentation
Traceability is very important in that a test result can be traced
from the time a sample is received in the lab to the result itself.
Baby Jade case
Applications in Local Cases
Baby Jade Case
A newborn baby was found abandoned in a trash container wrapped in
plastic.
Case was covered extensively by local media.
A woman came forward after the intense media coverage claiming to be
the mother.
Baby later died in a hospital.
A man claiming to be the father claimed the body of the baby.
Applications in Local Cases
What should have been done:
DNA tests should have been performed on the baby, alleged mother and
father to determine if they were really the parents of the child.
A result of maternity can be the basis for prosecution of the mother for
violation of the Child Protection Act and even infanticide as the baby
was not only left but there was intent to kill the baby by wrapping it in
plastic to suffocate it.
Baby Jade case
NEWS
Mom, who left Baby Jade in trash bin, charged with frustrated
infanticide
Published September 2, 2016 9:04pm
- See more at:
http://www.gmanetwork.com/news/story/579952/news/metro/
mom-who-left-baby-jade-in-trash-bin-charged-with-frustrated-
infanticide#sthash.Lvg0nMUT.dpuf
Applications in Local Cases
A result of paternity should have been the basis for releasing the
body of the baby to the man rather than just believing him.
In addition, if the mother claimed to have been raped which
resulted in the babys birth, the result can be the basis for a rape
case against the father.
Jee Ick Joo case
Applications in Local Cases
Korean Kidnapping Case
The body was cremated leaving no body or DNA to identify the victim.
A golf club set said to be the victims was found at the funeral parlor.
The Secretary of Justice surmises the golf set may have been planted
only.
Applications in Local Cases
What should have been done:
There should have been an attempt to extract trace
DNA from the golf set.
If DNA was found and a DNA profile was produced, it
should have been used to identify the victim by
comparing the DNA to the victims parents or children.
This would have established the identity of the victim.
Presence of the victims DNA would negate the claim
of the golf set being planted.
Presence of other persons DNA can lead to suspects
identity
 Hindrances to using DNA in
Forensics
Need to train police force in crime scene investigation
including DNA collection with adequate safeguards to
establish chain of custody and avoid contamination.
Cost of testing numerous samples from one crime scene
alone.
Establishing strict protocols to insure accuracy of results
and being able to present evidence in court.
Training of analysts for results interpretation especially if
DNA mixtures are found at the crime scene or in vaginal
swabs in rape cases
The Future of DNA Testing

From Genotype to Phenotype

Use of DNA markers to predict the appearance of the person (crime suspect
or victim)
May replace the artists sketch based on eyewitness accounts (which may be
erroneous due to faulty recollection of the eyewitness)
The Future of DNA Testing

In the latest International Society for Human Identification Meeting

held in Minneapolis, Minnesota, USA Sept. 2016, scientists have
shown convincing proof of the power of DNA to predict ethnicity, hair
and eye color as well as produce illustrations of the person tested
which were amazingly very close to the appearance of the subject.
Genotype: from this
Phenotype: to this
Thank you for your
attention!