Bangabandhu Sheikh Mujibur

Rahman Science & Technology

University, Gopalganj

An assignment on Application of DNA fingerprinting in paternity dispute

Course Title : Molecular markers and diagnostics

Course Code: BGE-413

Submission Date: 14-08-2021

Submitted By Submitted To
Rayhan Parvej Shovon Dr.Md. Sharafat Ali

ID NO: 15BGE015 Assistant Professor,

Dept. of
4th year 1st semester Biotechnology and Genetic
Engineering ,
Dept. of Biotechnology & Genetic
Engineering Bangabandhu
Sheikh Mujibur Rahman Science
Bangabandhu Sheikh Mujibur & Technology University ,
Rahman Science & Technology Gopalganj ,8100
University , Gopalganj ,8100

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Introduction:
DNA fingerprinting is proven to be quite useful in determining an individual's paternity. The forensic
DNA analysis is often used to discover criminal acts such as homicide and rape, but it is also utilized in
situations of contested paternity, determining the identity of a deceased person, and cases of baby
swapping. Any DNA carrying biological trace evidence can be tested using DNA typing. Because the DNA
molecule's makeup is basically constant from cell to cell, the DNA in blood is identical to that found in
other biological materials such as hair, sperm, skin, and bone marrow.

DNA fingerprinting has been incorporated to the normal work of contested paternity cases in India as a
strong tool for forensic inquiry. Because of the limitations or invariability of loci studied, the previous
traditional inquiry based on blood antigen systems such as variable blood groups and HLA Tissue Typing
was no longer employed in such delicate instances.(Medicine, n.d.)

Paternity, or the legal status of being a father, can be proven in a number of ways. Paternity is
frequently assumed when a child's parents are married. A lawsuit known as a "paternity action" must be
filed to ascertain if a man is the father of a kid born out of wedlock. Fatherhood can be proven in such a
case if the putative father confesses paternity. Blood, blood stain, and mouth swab can all be used to get
adequate DNA typing samples. DNA typing analyzes strands of genetic material between the kid and the
putative father. By comparing strands from different parts of the genetic material, accuracy rates of
99.9% can be achieved. An purported father can be ruled out with 100% confidence thanks to DNA
testing. The purpose of this study is to conduct an evidence analysis to help the court in establishing
physical facts in criminal and civil cases.

History of DNA Finger Printing:

Wyman and White first proposed the notion in 1980, based on the discovery of a polymorphic
DNA locus with a number of variable-length restriction fragments known as restriction
fragment length polymorphisms (RFLPs), which are specific regions where restriction enzymes
break the DNA. The publication “Hypervariable Minisatellite Regions In Human DNA,” authored
by Alec Jeffreys in 1985, is the first in the history of DNA. When Jeffreys and his colleagues were
looking at the human myoglobin gene, they came upon a section that consisted of a 33-base-
pair sequence that was duplicated four times. Because the amount of tandem repeats varies
both within and across loci, this tandem repeat was dubbed a minisatellite, and comparable
areas were dubbed hypervariable. Nakamura developed the term variable number of tandem
repeats (VNTR) in 1987 to characterize unique loci in which alleles are made up of tandem
repeats with different numbers of core units. A complicated ladder of DNA fragments is
identified when DNA is extracted, cleaved using a specific enzyme, and hybridized under low-
stringency conditions with a probe consisting of the core repeat. This profession looks to be
one-of-a-kind for each individual. Later on, different core repetitions were extracted and
utilized to create a variety of fingerprinting probes. This method was unknown at the time, but

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its promise was obvious. In 1985, DNA fingerprinting was used for the first time in a case of
paternity testing, which was essentially a maternity test, because paternal DNA was missing.

In this rare instance, a mother and her 13-year-old son were detained at the airport upon
arrival in England from Ghana because officials suspected he was not her son. A DNA
fingerprinting test on each of them proved that they were telling the truth. Later that year, DNA
fingerprinting was used for the first time in forensic identification in a case that wonderfully
demonstrates the potential of DNA evidence to link crime sites, dismiss suspects, and support
convictions. A guy was apprehended for reportedly raping two kids twice and then killing them.
A DNA fingerprinting test performed on a sample of sperm lef found at the crime site revealed
that a male was responsible for both crimes, although it was not the detained
suspect.(Equipment, 2014)

He was let go, and the true criminal was apprehended. This technology is still used today to
produce DNA profiles for each individual in order to solve crimes or test paternity.

What is DNA fingerprinting?

Simply put, DNA is a type of genetic material. Fingerprinting is a technology that allows people
to be identified based on their DNA's molecular features. Because the amount of bases and
repetitions inside a locus is unique to each individual, this approach employs VNRT. For
example, one person's genome may have the sequence gatagata, which repeats 10 times,
whereas another's genome may contain the identical pattern but only repeats 5 times. The
method is utilised in paternity testing and forensic investigations, as we've seen, but it may also
be applied in anthropological genetics, zoology, and botany, among other fields. Importantly,
the DNA Fingerprinting technology is extremely sensitive, which means it can yield data even
from half-decomposed biological material.(Genética et al., 2017)

Procedure to create a DNA fingerprinting:

Other molecular biology methods, such as polymerase chain reaction (PCR) and
electrophoresis, are utilized in these phases. The processes for creating a DNA fingerprinting
are as follows.

1. DNA is retrieved from any cell in the body's nucleus.

2. The restriction endonuclease enzyme (also known as the chemical knife) breaks down DNA
molecules into pieces. The VNTRs are also found in Te DNA fragments.

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3. In an agarose gel, Te fragments are sorted by size using gel electrophoresis.

Fig:01, Schematic illustration of procedure in DNA fingerprinting

4. Single-stranded DNA fragments are isolated and deposited onto a nylon membrane. Over the
nylon membrane, radioactive DNA probes with repeating base sequences corresponding to
potential VNTRs are poured. They will bind to single-stranded VNTRs in some cases. Southern
Blotting is a technique for hybridizing DNA using probes.

5. Extra probes are removed from the nylon membrane by washing it.

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6. The nylon membrane is subjected to an X-ray film to identify the locations where the
radioactive DNA probes have bonded to the DNA fragments. When X-ray film is processed,
these areas appear as dark bands. This is referred to as autoradiography.

7. The DNA fingerprints are shown by dark bars on X-ray film.

DNA Fingerprinting applications:

The DNA fingerprinting technique, which was invented by Alec Jeffreys, has been utilized in a
variety of scientific disciplines since its inception. In forensic investigations, it has aided in the
incarceration of offenders and the identification of victims of crimes, natural disasters, and
conflicts. This approach has been used to settle paternity cases. Furthermore, profile research
has been pushed by fields such as anthropological genetics, biology, and botany, among others,
in order to explain the origin and behavior of particular species. In the following paragraphs,
we'll go through how the approach has been used and evolved in the above-mentioned
domains.(Kingdom & States, n.d.)

paternity dispute:
DNA fingerprinting is a useful tool for determining the paternity of disputed offspring, for
example. Cases of baby exchanging, for example. This approach took the place of ABO blood
testing. antigen systems that can't prove paternity but can prove paternity To definitively rule
out an alleged father as a contender. Affliation orders are the source of disputed paternity. The
legality of divorce procedures has also been questioned. paternity can be established in
circumstances of inheritance, guardianship, and adoption. Maintenance, legitimacy, adultery,
or fornication are all words that come to mind while thinking of fornication. A DNA comparison
is done between progeny and prospective parents in parentage testing. Children inherit half of
their alleles from each parent, thus they should have a mix of alleles from both parents.(Gene &
Twins, 2016) The use of DNA profiles to identify whether an individual is the biological parent of
another individual is known as DNA paternity testing. Paternity testing is especially essential
when the father's rights and responsibilities are in question and the paternity of a child is in
question. Tests can also be used to establish whether or not someone is a biological
grandparent. Although genetic testing is the most accurate approach, there are other options,
such as ABO blood group typing, protein and enzyme analysis, or the use of human leukocyte
antigen antigens. Paternity testing now uses polymerase chain reaction (PCR) and restriction
fragment length polymorphism (RFLP). Paternity testing may now be done by a blood sample
while the woman is still pregnant.

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The most modern and accurate method for determining parentage is DNA testing. When the
alleged parent is not biologically connected to the kid, the result (called the "probability of
parentage") [failed verification] is 0%, and when the alleged parent is biologically linked to the
child, the likelihood of parentage is generally 99.99 percent. However, while virtually everyone
has a single set of genes, a small percentage of people, known as "chimeras," have at least two
sets of genes, which might lead to a false negative result if their reproductive tissue has a
different genetic make-up than the tissue collected for the test.(Gene & Twins, 2016)

DNA fingerprinting is proven to be quite useful in determining a person's paternity. A lawsuit

known as a "paternity action" must be filed to ascertain if a man is the father of a kid born out
of wedlock. DNA fingerprinting was first used in judicial cases, primarily to aid in the
investigation of crimes and the determination of paternity. It may also be used to find genetic
matches between tissue donors and recipients and to discover inherited genetic disorders.

Testing for paternity or maternity, whether for a kid or an adult:

A buccal or cheek swab is used to collect buccal (cheek) cells found on the inside of a person's
cheek for the DNA test. These swabs feature a cotton on synthetic tip and wooden or plastic
stick handles.

The collector massages the inside of a person's cheek to collect as many buccal cells as possible,
which are subsequently submitted to a lab for analysis. It would be necessary to collect samples
from the purported father or mother as well as the kid.

Prenatal paternity testing for a kid in the womb:

Prenatal paternity testing that is invasive:

Using techniques such as chorionic villus sampling or amniocentesis, it is possible to identify

who the biological father of the child is while the mother is still pregnant. Placental tissue is
obtained by chorionic villus sampling, which can be done transcervically or transabdominally.
Amniocentesis is a procedure that involves putting a needle into the pregnant mother's
abdominal wall to extract amniotic fluid. Because these techniques take a sample straight from
the fetus, they are quite precise; nevertheless, there is a tiny chance that the mother could
miscarry and lose the pregnancy as a result. Both CVS and Amnio need the expectant lady to
see a maternal fetal medicine expert who will execute the procedure.(Equipment, 2014)

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Non-invasive prenatal paternity testing:

The ability to identify the biological father while the woman is still pregnant has been made
possible thanks to advances in genetic testing. During pregnancy, the mother's blood contains a
tiny quantity of fetal DNA (cffDNA). This enables reliable fetal DNA paternity testing from a
blood draw during pregnancy with no danger of miscarriage. CffDNA may be detected as early
as 7 weeks gestation, according to studies, and the quantity of cffDNA rises as the pregnancy
develops.(Equipment, 2014)

DNA profiling:
In a criminal investigation, DNA fingerprinting is a laboratory method used to establish a
relationship between biological evidence and a suspect. A DNA sample from a crime scene and
a DNA sample from a suspect are compared. If the two DNA profiles match, the evidence was
obtained from that individual. If the two DNA profiles do not match, the evidence cannot have
originated with the suspect. Paternity can also be established via DNA fingerprinting.

There are various methods for analyzing DNA to establish if two samples are the same or
different. This is sometimes referred to as DNA fingerprinting. For example, two cloned pieces
of DNA can be studied in the laboratory to determine if they have portions in common, and
thus overlap with one another. In a different setting, such as a crime scene, DNA samples can
be collected and analyzed to determine if they match DNA samples obtained from suspects of
that crime. If two DNA samples have the same fingerprint, then there is a very high statistical
likelihood that they came from the same person. Such an approach can also be used to
establish paternity.(Medicine, n.d.)

Every somatic (nonreproductive) cell has the same DNA as an individual. Because sexual
reproduction combines the DNA of both parents to create a unique combination of genetic
material in a new cell, an individual's genetic material is derived in equal amounts from each
parent's genetic material; this genetic material is known as the individual's nuclear genome
because it is found in the nucleus.

Although DNA paternity tests can establish if one person is descended from the other by
comparing their DNA sequences, they are not yet 100 percent reliable. Specific sequences are
checked to determine if they were replicated verbatim from one individual's genome; if so, one
individual's genetic material may have been taken from the other (i.e. one is the parent of the
other). Mitochondria have their own genetic material, mitochondrial DNA, in addition to
nuclear DNA. Mitochondrial DNA is inherited only from the mother, with no scrambling.

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 Fig: 02, illustration of DNA finger printing

It is considerably easier to prove a link based on mitochondrial genome comparisons than it is

to prove a relationship based on nuclear genome comparisons. Testing the mitochondrial
genome, on the other hand, can only show if two people are connected through maternal lines
solely from a common ancestor and is thus of limited utility (i.e., it could not be used to test for
paternity).(Genética et al., 2017)

Because the Y chromosome is transmitted straight from father to son, it can be utilized to
determine paternity in a male kid.

The AABB regulates DNA paternity and familial connection testing in the United States,
however AABB certification is not needed. If the collection and processing of DNA samples

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follow a chain of custody, the findings are legally acceptable. Similarly, the SCC has standards
on DNA paternity and relationship testing in Canada, although this accreditation is not needed.

An example of DNA profiling used to identify a child's father (Ch).

A combination of various size DNA bands from both parents should be present in the child's
DNA sample. Person #1 is almost certainly the father in this scenario.

The International Society for Forensic Genetics' Paternity Testing Commission has taken on the
responsibility of developing bio-statistical recommendations in line with ISO/IEC 17025
standards. The Paternity Index, or PI, is a biostatistical estimate of paternity based on the
probability ratio concept. The guidelines give assistance on genetic hypothesis ideas and

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 mathematical problems required to create accurate PIs, as well as particular population
genetics difficulties.

Reverse paternity testing:

When the biological father of a person is unavailable, reverse paternity determination can be
used to determine who that person's biological father is. To produce a rough amalgamation,
the test utilizes STR alleles in the mother and her kid, other children and brothers of the
claimed father, and genetic laws to deduce the father's genetic constitution. When a direct
sample of the father's DNA is unavailable, this might be used to compare the father's DNA. This
test is seen on an episode of Solved being used to determine if a blood sample matches the
kidnapped victim.

Significance:
In a criminal investigation, DNA fingerprinting is a laboratory method used to establish a
relationship between biological evidence and a suspect. A DNA sample from a crime scene and
a DNA sample from a suspect are compared.

Because the DNA of every tissue from an individual has the same degree of polymorphism, DNA
fingerprinting is frequently employed in forensics. Paternity testing is based on DNA
fingerprinting since a kid inherits polymorphism from both parents. It may be used to
investigate population evolution and genetic diversity.

The precision of DNA profiling is maybe its most significant benefit. A DNA sample is collected
from each participant in the test – a kid, prospective father (and mother if she chooses to
participate) – using a simple, painless cheek swab (no blood or needles required – certainly a
benefit!).(Medicine, n.d.)

The main drawback of DNA fingerprinting is that it is not completely accurate. With this
technology, there are still issues about contamination, falsification, and chain of custody. Even
poor testing techniques might result in misleading positive or negative findings.

DNA fingerprinting is accurate 99.9% of the time.

 DNA fingerprinting is unobtrusive. ...
 DNA fingerprints have more than a criminal justice emphasis. ...
 DNA fingerprinting does not require a specific sample size. ...
 Data protection issues create additional storage and privacy issues. ...
 People are overly influenced by DNA evidence.

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 Some disadvantages, or risks, that come from genetic testing can include:
 Testing may increase your stress and anxiety.
 Results in some cases may return inconclusive or uncertain.
 Negative impact on family and personal relationships.
 You might not be eligible if you do not fit certain criteria required for testing.

One of the major discoveries of the late twentieth century, DNA fingerprinting, has transformed
forensic investigations. This study summarizes 30 years of development in forensic DNA
analysis, which aids in the conviction of offenders, the exoneration of wrongfully accused
individuals, and the identification of victims of crime, catastrophes, and conflict.

Conclusions:
Because the DNA fingerprinting technique allows identifying patterns in the known coding
region of genetic material that makes each individual unique, it has become an important tool for
scientific research. As a result, forensic investigations and parentage testing have found a tool to
convict criminals, identify victims, and resolve parentage disputes.

For the past thirteen years, we have successfully profiled and identified individuals, resulting in
enhanced rates of missing person identification using tests performed on the corpses of
unidentified people. By establishing a stable relationship between medical science and the
system of jurisdiction, the method was applied in cases of disputed parentage in civil cases,
which increased the importance of evidence and facilitated the court's awarding of just sentences,
which is critical for the future of children.

DNA profiling has transformed forensic genetics and is now frequently used in medical and legal
situations. DNA analysis is the most reliable method for conclusively excluding innocent people.
DNA testing has become the darling of the criminal and civil justice systems all over the world
as a result of all of these amazing uses.

References:

 Equipment, B. (2014). Application of DNA Fragment Analysis in

Disputed Paternity ( Maternity ) and Identification of the. 2818.
https://doi.org/10.5504/BBEQ.2012.0086
 Gene, N., & Twins, D. (2016). Ethical , Legal and Social Issues ( ELSI
) DNA : Statistical Analysis Allotypes , Immunoglobulin Biology /

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 DNA FORENSIC SCIENCES | DNA Profiling DEOXYRIBONUCLEIC
ACID PROFIL- ING | Overview. 2016–2018.
 Genética, H., Garcia, D., & Miño, K. (2017). NOTICIAS Y OPINIONES
/ NEWS AND VIEWS DNA fingerprinting. 477–480.
 Kingdom, U., & States, U. (n.d.). DNA paternity testing.
 Medicine, F. (n.d.). DNA Fingerprinting and Its Application in
Paternity Testing DNA basics.
 https://www.google.com/search?q=dna+fingerprinting+in+patern
ity+testing+significance&source=lmns&bih=664&biw=1536&hl=en
&sa=X&ved=2ahUKEwjd4MaQ6LfyAhUn5XMBHVIJAE8Q_AUoAHo
ECAEQAA
 https://commons.wikimedia.org/wiki/File:DNA_paternity_testing
_en.svg
 https://www.sciencedirect.com/topics/biochemistry-genetics-
and-molecular-biology/paternity-test/pdf
 Lecture and classes of Dr. Md Sharafat ali
 internet and associate books and references

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