Molecular diagnosis of diseases

✓ The structural and functional genomics

approaches have now pinpointed the
technical challenge in the exploration of
disease-related genes and the recognition of
their structural alterations or elucidation of
gene function.
✓ Various promising technologies and
diagnostic applications of structural
genomics are currently preparing a large
database of disease-genes, genetic alterations
etc., by mutation scanning and DNA chip
technology.
 Molecular diagnosis of diseases
✓ Further the functional genomics also
exploring the expression genetics
(hybridization-, PCR- and sequence-based
technologies), two-hybrid technology, next
generation sequencing with Bioinformatics
and computational biology.
✓ Advances in microarray “chip” technology
as microarrays have allowed the parallel
analysis of gene expression patterns of
thousands of genes simultaneously.
 Molecular diagnosis of diseases
✓ Further the functional genomics also
exploring the expression genetics
(hybridization-, PCR- and sequence-based
technologies), two-hybrid technology, next
generation sequencing with Bioinformatics
and computational biology.
✓ Advances in microarray “chip” technology
as microarrays have allowed the parallel
analysis of gene expression patterns of
thousands of genes simultaneously.
 Molecular diagnosis of diseases
✓ Sequence information collected from the
genomes of many individuals is leading to
the rapid discovery of single nucleotide
polymorphisms or SNPs.
✓ Further advances of genetic engineering have
also revolutionized immunoassay
biotechnology via engineering of antibody-
encoding genes and the phage display
technology.
✓ The Biotechnology plays an important role in
the development of diagnostic assays in
response to an outbreak or critical disease
response need.
 Molecular Diagnostic Techniques in
Identification and Characterization
✓ Correct and accurate identification of causative
agents like microbes in microbial diseases,
particular genetic sequences in genetic diseases
and protein levels are very essential for the
management of these patients, making specificity
and sensitivity important tools in diagnosis.
✓ Classical molecular techniques like normal PCR
and blotting although played satisfactory role in
diagnosis.
✓ However, currently molecular techniques like
gene and peptide sequencer, real-time PCR and
microarrays may detect more precisely and
specifically without consuming much time.
 DNA Paternity Test

✓ A DNA paternity test is nearly 100% accurate at

determining whether a man is another person’s
biological father. DNA tests can use cheek swabs
or blood tests. You must have the test done in a
medical setting if you need results for legal
reasons. Prenatal paternity tests can determine
fatherhood during pregnancy.
 DNA Paternity Test

✓ What is a DNA paternity test?

✓ Deoxyribonucleic acid, or DNA, is the genetic

material you inherit from your mother and father.
Paternity refers to fatherhood. A DNA paternity
test uses DNA, usually taken from a cheek swab,
to determine whether a man is the child’s
biological father.
 DNA Paternity Test

✓ How accurate is a DNA paternity test?

✓ DNA paternity tests are extremely accurate. A

test can show with 99.9% accuracy if a man isn’t
a person’s biological father.
 DNA Paternity Test

✓ How is a DNA paternity test performed?

✓ There are two equally accurate ways to test for

paternity:

✓ Blood tests: The potential father and child give

blood samples at a medical office. The facility sends
the samples to a lab for analysis.
✓ Cheek swabs: The potential father and child swab
the inside of their cheeks for buccal (cheek) cells.
You mail the cotton swab applicators to a
designated lab. If swabbing takes place in a medical
setting, the office sends the samples to a lab.
 DNA Paternity Test

✓ How is paternity confirmed?

✓ The lab runs a series of tests called DNA

sequencing. These tests look for genetic matches
between the potential father and child. A match
confirms paternity.
 DNA Paternity Test

✓ The laboratory analysis tests the DNA isolated from

cheek swabs to locate certain regions of
chromosomes that are known to vary in length
between individuals.
✓ Twenty Four of these sites are tested; each site is
called a “locus”, (“loci” – plural).
✓ Analysis of these 20 sites in a large population has
revealed many different sized versions associated
with each site. Versions of a DNA sequence or a
gene are called “alleles”.
 DNA Paternity Test

✓ The columns marked “allele” on the DNA test

report contain numbers indicating the two alleles
found at each locus (or one number if they are the
same size).

✓ If, for example, a child has two alleles that are

designated 12.1 and 18, and if the mother has alleles
12.1 and 16, then the child inherited the 12.1 allele
from the mother. The child has to have inherited the
18 allele from the father. The 18 allele is the
“obligate paternal allele.” Generally, the alleged
father must have this allele if he is the biological
father of the child.
 DNA Paternity Test

✓ 20 different loci are used as genetic markers in the

DNA tests, as well as one (Amelogenin) to confirm
the gender of the person providing the DNA sample.
✓ If the alleged father does not have the matching
allele at every tested locus, then he usually cannot
be the biological parent.
✓ If he does have it, then he could be the father. A
“relationship index” (called the “Direct Index” in
the report) for each locus is calculated based on
information including the portion of the male
population that has the obligate paternal allele at
that locus.
✓ This index is reported for each DNA locus.
 DNA Paternity Test

✓ A combined relationship (or “Direct”) index for all of the

tested alleles is then calculated and appears below the
chart. This number is used to calculate the “probability of
relationship,” which is the percentage likelihood that a
man with the alleles of the alleged father is the biological
parent of the child, as compared to an untested, unrelated
man of the same race.
✓ If the DNA of the alleged father is consistent (to a degree
of mathematical certainty) with that of the child, then the
report will conclude that the alleged father cannot be
excluded as the biological father of the child. If the DNA
is not consistent however, it will conclude that the alleged
father can be excluded as the biological father of the child
 DNA Paternity Test

✓ Each locus used in DNA testing is composed of a variable

number of repeating short sequences of the DNA bases. An
example of a natural DNA sequence having such simple short
repeats would be ACGACGACGACG, i.e. (ACG)4.
However, each allele can have a different number of total
repeats, for example (ACG)3 or (ACG)5 instead of (ACG)4,
giving rise to a different fragment length when the DNA is
purified and amplified in the test tube.
✓ In this example, (ACG)3, (ACG)4 and (ACG)5 would encode
variant alleles with building block lengths of 3 x 3, 4 x 3, and
5 x 3, that is to say 9, 12, and 15 building block lengths.
Since DNA analysis machines detect changes in length of just
one unit, these differences in length are easily distinguished.
DNA Paternity Test