GEN E S TR UCTU RE,

EXPRESSI ON AN D MUTATI ON
 SIGNIFICANCE

• GENETIC ISSUES ARISE BEFORE, DURING

AND AFTER PREGNANCY
• POSITIVE FAMILY HISTORY OF DISEASES
• AMNIOCENTESIS OR CHORIONIC VILLUS
SAMPLING
• WORK-UP FOR INFERTILITY
• DISORDERS OF SEXUAL DEVELOPMENT
• GYNECOLOGIC MALIGNANCIES
 Chromosome analysis
can be performed on

• Peripheral blood – for

analysis of fetal,
neonatal, juvenile, or
adult blood
• Bone marrow - for
malignant studies
• Tissue – for malignant
studies
Chromosome analysis
can be performed on

• Fetal tissue – for identification of

reasons for fetal demise and for
tissue culture
• Amniotic Fluid
• Chorionic Villi
• Umbilical Cord Blood
Chromosome analysis is indicated in
the following cases

Individuals with
• suspected classic chromosomal syndrome
• multiple congenital anomalies
• dysmorphic features
• failure to thrive
• ambiguous genitalia
• abnormalities of sexual development
• amenorrhea
• short stature
• mental retardation of undetermined etiology
• developmental delay
Couples with
• two or more miscarriages
• Infertility
• Advance age

Family members
• both parents of a child with structural
chromosome rearrangement, deletion, or
duplication
• at risk of having a chromosome
rearrangement

Pregnancy
• abortuses
• stillbirth
 Sample
Collection

Peripheral Blood Sterile sodium or lithium heparinized tube (green top

vacutainer) or sterile tube with 0.05ml preservative free
heparin.
Ensure that sample is no clotted
Specimen
Culture

Blood:
Children/adults - 8 drops of whole
blood/ 5 ml culture
media
Infants - 4-5 drops/ 5 ml
culture media
Bone marrow:
4 –5 drops into 8 ml
T/C flask of media 2 flasks
Per sample
 Specimen
Incubation

Incubate (37oC)
Blood = 72 Hours
Bone marrow = overnight
 Harvest
Cell cycle

• Metaphase arrest
• Hypotonic swelling
• Lysis by acetic acid
• Fixation
Metaphase
Arrest

Metaphase arrest
(Colchicine/ Colcemid)
Increasing Cell
Volume by KCl

Hypotonic shock
The hypotonic solutions:
.075M KCL dilute balance salt
solution, sodium citrate, 20%
diluted serum in H2O
Lysis by Acetic
Acid
 Fixation

Fixation
Methanol & Glacial
acetic acid (3:1)
Slide Preparation

Slides
 Chromosome
Banding
C-banding stains centromeres.

R-banding is the reverse of C-banding and stains non-

centromeric regions in preference to centromeres.

G-banding is obtained with Giemsa stain. It yields a series

of lightly and darkly stained bands (right).

Q-banding is a fluorescent pattern obtained using

quinacrine for staining. The pattern of bands is very
similar to that seen in G-banding.

T-banding produce specific staining of the telometric regions of the

chromosomes
GTG-banding

Chromosomes are G-banded to facilitate the identification

of structural abnormalities. Slides are dehydrated, treated
with the enzyme trypsin, and then stained.
Screening and
Analysis
Chromosomes are described with
the following categories:

Metacentric: centromere is median or near median chromosome has two

well defined arms with a length ratio varying from 1:1 to 2.5:1

Acrocentric: centromere is close to one end of the chromosome one arm is

substantially smaller than the other and the arm
ratio ranges from 3:1 to 10:1
Telocentric: centromere is a strictly terminal entity and the chromosome is
one armed
Capturing and
Analysis
Normal Male
Karyotype
Normal Female
Karyotype
DNA

• A molecule made up of linear sequence of

nucleotides intertwined as a double helix.

• Within cells, DNA is organized into structures

called chromosomes.

• -The main role of DNA molecules is the long-

term storage of information
• What are
Chromosomes?
Chromosomes are tiny
string-like structures in
cells of the body. They
contain the estimated
30,000 to 35,000
human gene pairs that
determine traits like
eye and hair color, as
well as direct the
growth and development
of every part of our
physical and biochemical
systems.
• Each person
normally has 23
pairs of
chromosomes, or
46 in all. We
normally inherit
one chromosome
per pair from our
mother and one
from our father.
Backbone:

• - Phosphate
– Pentose sugar
(deoxyribose)
– Nitrogen bases
 Bases in a DNA
molecule:
• Purines
Adenine and Guanine

• Pyrimidines

Thymine & Cytosine

 Gen e

• Biologic unit of heredity

• Located at a definite position (locus) on a
particular chromosome.
• consist of a long strand of DNA that contains a
promoter, coding and non-coding sequence.
• promoter- controls the activity of a gene,
• . Coding sequence - determines what the gene
produces,
• non-coding sequence - regulate the conditions of
gene expression.
 Gene

• When a gene is active, the coding and

non-coding sequence is copied in a
process called transcription, producing an
RNA copy of the gene's information
 Messenger
Ribonucl ei c
Ac id (mRN A)
• is a molecule of RNA encoding a chemical "blueprint" for
a protein product.mRNA is transcribed from a DNA
template, and carries coding information to the sites of
protein synthesis: the ribosomes.
• In mRNA as in DNA, genetic information is encoded in
the sequence of four nucleotides arranged into codons of
three bases each. Each codon encodes for a specific
amino acid, except the stop codons that terminate
protein synthesis. This process requires two other types
of RNA:
• Transfer RNA (tRNA) mediates recognition of the codon
and provides the corresponding amino acid,
• while Ribosomal RNA (rRNA) is the central component
of the ribosome's protein manufacturing machinery.
Tr anscri ptio n

Transcription is the synthesis of mRNA from a DNA

template. During transcription, RNA polymerase makes a
copy of a gene from the DNA to mRNA as needed.
Ste ps i n tra nsc rip tion

• DNA unwinds.
• RNA polymerase recognizes a specific base
sequence in the DNA called a promoter and
binds to it. The promoter identifies the start of a
gene, which strand is to be copied, and the
direction that it is to be copied.
• Complementary bases are assembled (U
instead of T).
• A termination code in the DNA indicates where
transcription will stop.
• The mRNA produced is called a mRNA
transcript.
 Pr ocessin g th e
mRNA Tr anscrip t
• the newly-formed mRNA transcript (also called
heterogenous nuclear RNA or hnRNA) must be
further modified before it can be used.
• A cap is added to the 5’ end and a poly-A tail
(150 to 200 Adenines) is added to the 3’end of
the molecule.
• The newly-formed mRNA has regions that do
not contain a genetic message. These regions
are called introns and must be removed. Their
function is unknown.
• The remaining portions of mRNA are called
exons. They are spliced together to form a
mature mRNA transcript.
Processing the
mRNA
Transcript
 Translatio n

• Translation is the process where ribosomes

synthesize proteins using the mature mRNA
transcript produced during transcription.
• During translation, the mRNA transported to
the cytoplasm is "de-coded" or "translated" to
produce the correct order of amino acids in a
protein.
• tRNA = transfer RNA; small RNA molecules that
carry a specific amino acid at one end and an
anticodon region that recognizes and binds
mRNA at the other end. The tRNA that binds to
that mRNA codon determines what amino acid is
added to a protein chain.
The steps of
transl ati on:
• 1. Initiation: mRNA enters the cytoplasm and becomes associated
with ribosomes (rRNA + proteins).
• tRNAs, each carrying a specific amino acid, pair up with the mRNA
codons inside the ribosomes. Base pairing (A-U, G-C) between
mRNA codons and tRNA anticodons determines the order of
amino acids in a protein.

• 2. Elongation: addition of amino acids one-by-one:

As the ribosome moves along the mRNA, the tRNA transfers its
amino acid to the growing protein chain, producing the protein -
codon by codon!

• 3. Termination: when the ribosomes hits a stop codon - UAA,

UGA, or UAG - the ribosome falls apart!
The same mRNA may be used hundreds of times during translation
by many ribosomes before it is degraded (broken down) by the cell.
 Steps in
Translocation
 Mutation

Gene Mutation-
• Occurs as a result of environmental damage to
DNA, through errors during DNA replication or
repair.
• Reserved for new changes in the genetic code
that lead to altered function and clinical
consequences.
Point Mutations- results in amino acid
substitution, leading to different products with
altered functions. E.g. sickle cell anemia
Mol ecul ar Tool s and
Di agnosi s in Human
Geneti cs

1. PCR
2. RE/ RFLP
 Mol ecul ar Tool s and
Di agnosi s in Human
Geneti cs

Polymerase Chain Reaction- a common

method of creating copies of specific
fragments of DNA. PCR rapidly amplifies a
single DNA molecule into many billions of
molecules.
 Rest ric tio n En do nuc lea ses
an d R estric tion F ragm en t
Le ngt h Po lymo rph isms

• are enzymes that recognize and cut specific nucleotide

sequences in the double stranded DNA molecule
• the sites of their action are known as restriction sites
• The restriction fragment length polymorphisms can be used
to follow the transmission of a gene in a family.
 Mi cro arra y
DN A An alysi s
• Permits the expression and analysis of
thousands of genes simultaneously.
• It was used to understand the molecular
basis of cancer and the biological behavior
of tumors.
• This powerful tool can provide a molecular
fingerprint of an individuals disease and is
referred to as gene expression
profiling(GEP)
Genetic Testing:
Direct and
In dir ect Me thods

Direct Method
• Direct testing for the actual mutation in an
affected individual to confirm the clinical
diagnosis or prenatal diagnosis would be
possible by obtaining DNA from the
subject.
 Indi rect
Method

• when direct testing is not possible

• - DNA markers located to the presumptive
disease-causing gene/mutation are used
as road maps to identify the travel or
passage of the gene from an affected
parent to an at-risk offspring.
 Indi rect
Method
• This requires that the affected individual
has markers that are informative, in other
words, unique or distinctive from markers
of the non affected individual.
• Multiple family members, both affected
and unaffected must have DNA available
for analysis in order for this approach to
be informative.
• The markers are often RFLPS.
 Mol ecul ar
Cyt oge net ics
• powerful tool to analyze chromosome abnormalities that
are not visible using traditional karyotyping and
microscopy.
• most widely used procedure is fluorescent in situ
hybridization(FISH)
• FISH takes advantage of the complementary nature of
DNA
• Denatured DNA sequences labeled with a fluorescent
dye are hybridized onto denatured chromosomes that
have been immobilized onto a slide. The chromosomes
are then viewed with a wavelength of light that excites
the fluorescent dye.
 Mol ecul ar
Cytogenetics
• FISH is commonly used to screen for
chromosome anueploidy in amniotic fluid cells in
prenatal diagnosis
• Powerful tool to confirm or diagnose syndromes
that are due to microdeletions of segments of
chromosomal material
• Used to identify the actual physical location of a
gene or to order a serias of DNA sequences or
genes on a chromosome
Comparative Genome Hybridization- used to
measure differences in copy number or dosage
of a particular chromosome segment.
• its widest application is in the study of a gene
dosage in normal and cancer cell lines.
Spectral Karyotyping- uses the FISH principle to
visualize all 24 chromosomes by “painting” with
chromosome-specific probes in different colors
simultaneously.
 Patterns of
Inheritance
Pedigree
= graphic representation of family
history data that assists in mining the
transmission pattern of the gene

= the pattern of transmission and the

constellation of the characteristics of
the affected individuals in the
pedigree confirms the diagnosis
 As a review….

• A recessive trait is one that is "obscured" by

dominant traits, but is expressed when two
recessive genes are present. When
symbolizing recessive traits, lower-case letters
are used (aa).
• A dominant trait is one that is expressed even
in the presence of other genes for the same
trait. When symbolizing dominant traits, a
capital letter is used (AA or Aa).
 AUTOSOMAL
DOMINANT

• Only one of the mutated gene is

required for expression of the
trait
• Individual is said to be
heterozygous for the trait
 General
Characteristics
– Every affected individual has an
affected parent.
– If reproductively fit, the affected person
has a 50% chance in transmitting the
gene with each pregnancy.
– The sexes are affected equally.
– There is a father to son transmission.
– An individual who does not carry the
mutation will have no risk of
transmission to his or her offspring.
Three additional
properties
• variable expressivity= severity of
phenotype in individuals who have
the mutation
• penetrance= probability that a gene
will have any clinical manifestation
at all in a person known to have the
mutation
• new mutations
AUTOSOMAL
RECESSIVE
– rare; require the affected individual to
have two copies of the mutant allele
(homozygous) in order to manifest the
condition
 GENERAL
CHARACTERISTICS

• The characteristic will occur equally in both sexes.

• for an offspring to be at risk, both parents must have at least
one copy of the mutation
• If both parents are heterozygous (carrier) for the condition,
25% of the offspring will be homozygous for the mutation and
manifest the condition, 50% will be carriers and unaffected.
The remaining 25% will not have inherited the mutation at all,
will be unaffected, and will not be at risk of transmitting the
mutation to any offspring.
• consanguinity is often present in families demonstrating rare
autosomal recessive conditions.
• If the disease is relatively rare, it will be clustered among the
siblings, and will not be seen among other family members
such as ancestors, cousins, aunts and uncles.
X-LINKED TRAIT

-diseases caused by genes on the x

chromosome
– Most are recessive
– Expression of genes located on the x
chromosome demonstrate a unique
characteristic known as DOSAGE
COMPENSATION
– Achievement of dosage compensation
is through the principles of X
inactivation, also known as the LYON
HYPOTHESIS
LYON HYPOTHESIS

• One x chromosome in each cell is

randomly inactivated in the early female
embryo (soon after fertilization).
• The inactivation process is random; either
the paternally or maternally derived x
chromosome is chosen. The female is
thus a mosaic for genes located on the x
chromosome.
• All descendants of the cell will have the
same inactive X chromosome.
X-LINKED DOMINANT
INHERITANCE

– All heterozygotes, both male and

female, manifest the condition
– Affected males never have affected
sons, and all daughters of affected
males are affected.
 Atypical Patterns of
Inheritance

• TRINUCLEOTIDE-REPEAT DISORDERS: UNSTABLE

MUTATIONS
• A new class of genetic conditions in which the
gene mutation was dynamic and would change
with different affected individuals within a family
• Most common group of disorders
• Unstable in that they tend to expand as the gene
is passed on from generation to generation----
progressively earlier onset or more severe
manifestations of disease with each successive
generation---- anticipation (generally
characteristic of genetic conditions caused by
unstable repeats)
• Molecular mechanism: misalignment at the time
of meiosis
• FRAGILE X SYNDROME

• Most common heritable form of moderate

mental retardation
• Gene is located on X chromosome Xq27.3
• FMR1 (fragile X mental retardation 1)
--- triplet expansion blocks normal function of
the
FMR1 gene
• Individuals with an intermediate number
of copies (52-200)= PREMUTATION
CARRIERS= unaffected
• GENOMIC IMPRINTING AND UNIPARENTAL
DISOTOMY

• Differential activation or expression

of genes depending on the parent of
origin
• A group of diseases in which the
parent of origin of a gene or
chromosome plays a role in the
phenotype of the affected individual
 EXAMPLES

PRADER-WILLI SYNDROME
• Characterized by obesity, hyperphagia,
small hands and feet, hypogonadism, and
mental retardation

ANGELMAN SYNDROME
• Due to the absence of a paternal
contribution of the genes located at 15q11-
q13
• Flattened back of the head.
Deep-set eyes.
Wide, ever-smiling mouth.
Prominent jaw and widely spaced teeth.
Lightly pigmented hair, skin and eyes.
• GERMLINE MOSAICISM

• Presence of two or more genetically

different cell lines in the same
individual or tissue derived from a
single zygote
• Mutation is present in only one parent
and arose from embryogenesis in all
or some of the germ cells but few or
none of the somatic cells of the
embryo
• OSTEOGENESIS IMPERFECTA TYPE
II (LETHAL FORM)

• At the molecular level, the

mutation causing the
condition is dominant, that is,
only one copy of the abnormal
gene is necessary to cause
this perinatal lethal condition
 MITOCHONDRIAL
INHERITANCE- MATERNAL
INHERITANCE

• Growing number of conditions due to abnormalities of

mitochondria
• Do not follow the typical mendelian pattern of inheritance

MITOCHONDRIAL DNA (mtDNA)

• MITOCHONDRIAL DISEASES CAN BE CAUSED BY MUTATIONS IN
THE mtDNAOR IN THE NUCLEAR GENES THAT CODE FOR
COMPONENTS OF THE OXPHOS SYSTEM.

LEBER’S HEREDITARY OPTIC NEUROPATHY

• Rapid, bilateral loss of central vision occurs
MULTIFACTORIAL
INHERITANCE

• traits or characteristics produced by

the action of several genes, with or
without the interplay of
environmental factors.
-Cleft lip with or without cleft palate
-Open neural tube defect
-Cardiac defects
CHROMOSOME
ABNORMALITIES
• Numerical chromosomal abnormalities
1. aneuploidy- refers to an extra chromosome,
such as in Trisomy 21 (Down Syndrome),
Monosomy X (Turner Syndrome)
2. Polyploidy- refers to numerical chromosome
abnormalities in which there is addition of an
entire complement of haploid chromosome
such as triploidy, in which three haploid sets
occur (XXY or XYY).
• Numerical or aneuploid
chromosome abnormalities
invade either autosomes or sex
chromosomes. Most occur as
the result of nondisjunction
during meiosis or mitosis in
which homologous
chromosomes fails to disjoin
• Molecular structures for the
parent of origin have identified
that the majority of autosomal
aneuploidies result from non
disjunctional error in maternal
meiosis I.
• The majority of trisomic
conceptions are nonviable,
autosomal trisomies have been
seen in abortus material in all 21,
18, 13, and 22 result in live
chromosomes 1 and 17. However,
trisomies births and are associated
with advance maternal age
 Tris omy 1 3
(P atau Syndrome

• occurs in approximately 1/10,000

live births.
• Characterized by gross multiple
structural defecits:
procencephaly, cleft lip/palate,
cardiac defects, and polydactyly.
 Tris omy 1 8
(Ed ward
Syndrome)

• 1/6,000 of live births

• associated with prenatal
growth restriction, rocker
bottom feet, cardiac and
renal defects.
 Tri so my 21
(Do wn
Sy ndrome )
• most common viable autosomal
trisomy and has an incidence of
1/800 of live births.
*The majority (95 %) of individuals with
Down syndrome have trisomy 21
because of maternal disjunction.
• However, about 2% to 3% of
individuals with clinical Down
syndrome have structural
rearrangement (robertsonian
Translocation) and another
1 % to 3 % is mosaic for
trisomy 21.
 Tri so my 22

• has been seen in a few live born

individuals and is associated with
severe neurologic impairment.
*monosomic states involving
autosomes are extremely rare
and generally lethal.
Sex Chromosome
aneuploidy usually
occurs in the trisomic
states.
Monosom y Y

• has never been seen

in clinical situation or
even in abortus
Monosom y X
(45 X, Tur ner
Sy ndrome )
• typical finding, However, because
most 45 X conceptions are lethal, the
actual incidence of live births is
about 1/5,000.
• characterized by lymphedema,
hypotonia and webbed neck.
• girls with Turner syndrome have short
stature, broad chest with wide spaced
nipples, cubitus valgus ( widened
carrying angle of the arms), gonodal
dysgenesis resulting in lack of
secondary characteristics, amenorrhea,
and infertility.
• others features include congenital heart
disease (COA, most common), kidney
disease, and hypertension in later life.
• Intelligence is normal
although spatial perception
abnormalities are common.
• * Hormonal supplementation
during puberty allows girls to
develop secondary
Characteristics.
• the 45 X karyotype occurs
paternal non disjunction
• and is not associated with
advanced maternal or paternal
age.
• There is no recurrence for 45 X
• Another 30 to 40 % of individuals are
mosaic for the 45 X cell line and
another cell line (usually 46 XX)
because of postzygotic
nondisjunction during mitosis.
• Females who are
mosaic with a 45 X or
46 XY karyotype are at
an increased risk for
gonadoblastoma.
• Other trisomies involving the
sex chromosomes are seen in
47 XXX, 47 XXY (Klinefelter
Syndrome ), 47 XYY
karyotypes.
 47 XXX

• -1/1000 live births

• - due to maternal nondisjunctiuon associated
with increasing maternal age.
• - most women are phenotypically normal with
exception of possible mild developmental delay:
fertility is normal and there may be a slightly
increased risk for offspring with aneuploidy
involving the sex chromosomes and
autosomes.
 47 XXY
( kl inef elter
Sy ndrome )
• -1/1000 live male births
• -is a common sex
chromosome abnormality
associated with advanced
maternal age.
• Clinical features include tall
gynecoid stature, gynecomastia
(with an increased risk for breast
cancer) and testicular atrophy.
• -mental retardation is not a typical
feature, but affected individuals
may have IQ scores that are
lower than those of their siblings.
 47 XYY

• - nondisjunction during spermatogenesis

involving the Y chromosome.
• taller than average,but are otherwise
phenotypically normal
• contrary to previous and outdated observational
studies, this sex chromosomes aneuploidy is
not associated with violent crime. However,
behavioral problems such as attention deficit
disorder may be observed.
B . Str uct ur al chr omosomal
abnor mal ities
• Chromosomes breaks and
rearrangements may lead to no
obvious phenotypic
consequences (genetically
balanced), loss or gain of
chromosomal material
(genetically imbalanced) that
produce abnormalities, or
abnormalities due to
interruption of a critical gene at
 STRUCTURAL
CHROMOSOMAL
ABNORMALITIES

DELETION RING CHROMOSOME

INVERSION
ISOCHROMOSOM
E
DUPLICATION
TRANSLOCATION
INSERTION
 Ty pes:

• translocations (reciprocal and

robertsonian)
• insertions
• inversions
• isochromosomes
• duplications
• deletions
 Balanced
recip rocal
tr anslo cations
• translocations occur as a result
of a mutual and physical
exchange of chromosomes
material between non
homologous chromosomes.
• found in about 1/11000
newborns.
* The carrier of reciprocal
balanced translocations is
usually phenotypically normal.
However, the carrier is at an
increased risk for producing
offspring who are
chromosomally abnormal.
• In meiosis, the two pairs of
nonhomologous
chromosomes involved in the
translocation resolve their
pairing difficulties by forming a
quadriradial.
• Most reciprocal translocations are
unique to a family, and, consequently,
the reproductive fitness of the carrier
depends on the carrier’s sex and
nature of the translocations.
• In general, the recurrence for
unbalanced conception is 3% to 5%
for male carriers and 10% to 15% for
female carriers.
Ro bertsoni an
tr ansl ocati on

• structural rearrangement between the

acrocentric chromosomes:
chromosome pairs 13, 14, 15, and 22.
• in this structural rearrangement, the
short arms (p arm) of two
nonhomologous chromosomes are
lost, and the long arms fuse at the
centromere, forming a single
chromosome structure.
• The phenotypically normal carrier
of a robertsonian translocation
has a 45 chromosomes in each
cell because the two acrocentric
chromosomes involved in the
translocation have formed into
one chromosome structure.
• This person is genetically balanced,
that is he or she has two copies of
each chromosome. However, the
gametes are at risk to be unbalanced.
• As in reciprocal translocations, the
chromosomes involved in the
rearrangement resolve the pairing of
homologous segments at meiosis by
forming
Inversions
-occur when two breaks occur on a
chromosomes followed by 180o turn of the
segment and reinsertion at its original
breakpoints.
• Pericentric inversion-if the centromere is
included in the inverted segment
• Paracentric inversion- if the centromere is
not involved.
Isochromoso
mes
• occur as a result of the chromosome
dividing along the horizontal axis
rather than the longitudinal axis at the
centromere.
• the result is a chromosome that has
two copies of one arm and no copies
of the other.
* isochromosomes involving the
autosomes are generally lethal
because the resultant conception
will be both trisomic and
monosomic for genetic
information. However
ischromosomes involving the long
arm of the X chromosomes (iso Xq
) is compatible with life.
Deletions and
Duplications

• arise from unequal crossing

over at meiosis, or from
crossing over during pairing
of inversion or
translocations.
• Terminal deletions- break
resulting in loss of chromosomes
material at the tip.
• Interstitial deletions- a loss of
chromosome material between
two breaks within a chromosome.
Cri-du-chat (5p)
syndrome

• microcephaly, profound
mental retardation, growth
retardation, a unique facial
appearance, and a
distinctive”cat like” cry.
*most cases occur as a de novo
chromosome deletions of the
tip of the short arm of
chromosome 5, but
approximately 10% to 15%
arise as a consequence of
parental reciprocal
translocation chromosome 5
and another chromosome.
Mi crod eleti on/
dupl ication or
Conti guous
Gene
Syndr ome
• syndromes in which the involved
chromosome region are submicroscopic
and so small that molecular cytogenetic
technique such as fluorescent in situ
hybridization is necessary to localized the
affected region.
• The phenotypes of these conditions are
due to absence ( or duplication) of
multiple contiguous genes within the
involved region.
Chromosome Abnormalities
and Pregnancy Outcome
• incidence and types of
chromosome
abnormalities differ
between spontaneous
abortions, stillbirths, and
live births.
• chromosome and lethal genetic
abnormalities play a major role in
early losses. 30 to 60 % of first
trimester abortuses are found to
have a chromosome abnormality
of which approximately 50% are
due to autosomal trisomies.
• certain chromosomes are more
commonly involved than the
others; for example, trisomy 16
accounts for one third of trisomic
abortuses.autosomal monosomies
are extremely rare,accounting for
less than 1% of chromosomally
abnormal fetus. Triploidy accounts
for 16% of spontaneous abortions.
• Turner Syndrome due to 45 X
by far the most common
chromosome abnormality,
accounting for 20% of
spontaneous abortions that are
chromosomally abnormal.
Molar Gestation
Partial moles (associated with
triploidy)

• found in 16% of abortuses.

• they occur as a consequence of
errors in meiosis or from double
fertilization of a single ovum.
• clinical features depend on the
parent of origin of the extra set of
chromosomes. In 2/3 of cases,
there is one maternal set and two
paternal set sets of chromosomes,
resulting in poor embryonic
development but a large hydropic
placenta, the partial mole.
Conversely there is an
extra maternal set, an
anomalous and growth-
restricted fetus and a
small fibrotic placenta
are seen.
Complete
mole
• diploid with a 46 XX karyotype.
• paternally derived
• arise through the fertilization of
an unucleated oocyte and
subsequent duplication of haploid
chromosome complement (23 X)
of the sperm.
* the absence of maternal
contribution results in
development of abundant,
hydropic trophoblastic tissue,
and no recognizable
embryonic tissue.
 Down
Syndr ome

47,XY,+21
Ed wa rd’s
Syndrome

47,XY,+18
Translocation

46,XY,t(1;17)
 BCR/ABL
Metaphase
• A metaphase cell
positive for the bcr/abl
rearrangement using
FISH. The chromosomes
can be seen in blue. The
chromosome that is
labeled with green and
red spots (up left) is the
one where the wrong
rearrangement is present
 BCR/ABL
Interphase
• Interphase cells
positive for a
chromosomal t(9;22)
rearrangement.
 NUMERICAL
CHROMOSOMAL
ABNORMALITIES
• Polyploidy is a state in which the number of
sets of chromosomes exceeds the diploid
number (2n) by a multiple of n
• Triploidy - A complete extra set of
chromosomes raises the total number to 69.
This usually arises from:
- Fertilization by two sperm (dispermy)
- Failure of one of the maturation
divisions of either the egg or the
sperm, producing a diploid gamete
 TRIPLOIDY
• A triploid fetus (which usually miscarries) would
be 69,XXY (most common), 69,XXX or 69,XYY
depending on the origin of the extra set of
chromosomes
 TETRAPLOIDY

• Tetraploidy - Four times the haploid

number is usually due to failure to
complete the first zygotic division

• Tetraploid cells are also a normal feature

of regenerating liver and other tissues
 POLYPLOID

• A proportion of cells that occurs normally

in human bone marrow, as
megakaryocytes usually have 8-16 times
the haploid number
BIOCHEMICAL
GENETICS
BIOCHEMICAL
GENETICS

• RENDERS SERVICES FOR THE

DIAGNOSIS AND CLINICAL
MANAGEMENT OF PATIENTS WITH
INBORN ERRORS OF METABOLISM
(IEM)
• PROVIDES KEY INVESTIGATIONS FOR
VARIOUS METABOLIC DISORDERS
INBORN ERRORS
OF METABOLISM

• DISORDERS THAT ARISE FROM A

DEFECT IN A METABOLIC PATHWAY,
WHICH COULD BE DUE TO EITHER AN
ENZYME OR AN ABNORMALITY OF A
TRANSPORT PROCESS
COMMON SIGNS
AND SYMPTOMS

• ACUTE ENCEPHALOPATHY
• CHRONIC ENCEPHALOPATHY
– MENTAL RETARDATION
– DEVELOPMENTAL DELAY
– SEIZURES, ETC
• MYOPATHY
• DIFFUSE LIVER DISEASE
• RENALTUBULAR DISEASE
 TYPES OF IEM

• DISORDERS OF PROTEIN METABOLISM

– (AMINO ACIDOPATHIES, ORGANIC ACIDOPATHIES,
UREA CYCLE DEFECTS)
• DISORDERS OF CARBOHYDRATE METABOLISM
– (CHO INTOLERANCE DISORDERS, GLYCOGEN
STORAGE DISORDERS, DISORDERS OF
GLUCONEOGENESIS AND GLYCOGENOLYSIS)
• LYSOSOMAL STORAGE DISORDERS
• FATTY ACID OXIDATION DEFECTS
• MITOCHONDRIAL DISORDERS
• PEROXISOMAL DISORDERS
 LOCALLY
AVAILABLE TEST
• COMPREHENSIVE URINE METABOLIC
PROFILE (URINE METABOLIC SCREEN
AND ORGANIC ACID ANALYSIS)
• URINE METABOLIC SCREEN (AMINO
ACID PROFILE, METHYLMALONIC
ACID SCREEN)
• PLASMA AMINOACID QUANTITATIVE
ANALYSIS BY HIGH PERFORMANCE
LIQUID CHROMATOGRAPHY(HPLC)
 LOCALLY
AVAILABLE TEST

• CSF AMINOACID QUANTITATIVE

ANALYSIS BY HPLC
• URINE ORGANIC ACID ANALYSIS FOR
GAS CHROMATOGRAPHY-MASS
SPECTROPHOTOMETRY(GCMS)
• EXPANDED NEWBORN SCREENING
PRIMARY REASONS
FOR REQUESTING
GENETIC
METABOLIC TEST
• CHRONIC • HEMATOLOGIC
ENCEPHALOPATHY SYMPTOMS
• ACUTE • ENDOCRINE
ENCEPHALOPATHY
• METABOLIC ACIDOSIS
• PSYCHIATRIC
• BIOCHEMICAL
SYMPTOMS
ABNORMALITIES
• MOVEMENT DISODERS
• DYSMORPHIC
• STROKE
FEATURES
• MYOPATHY • OTHER VISCERAL
• OCULAR SYMPTOMS SYMPTOMS
• CARDIAC ARRYTHMIAS
• HEPATIC SYMPTOMS
MOLECULAR
GENETICS
 FLUORESCENT IN
SITU
HYBRIDIZATION
(FISH)
• A technique that can be used to detect and
localize the presence or absence of specific
DNA sequences on chromosomes. It uses
fluorescent probes that bind to only those parts
of the chromosome with which they show a
high degree of sequence similarity.
Fluorescence microscopy can be used to find
out where the fluorescent probe bound to the
chromosome. FISH is often used for finding
specific features in DNA. These features can be
used in genetic counseling, medicine, and
species identification.
FISH Principle
 Medical
appl ications
• In medicine, FISH can be used to form a
diagnosis, to evaluate prognosis, or to evaluate
remission of a disease, such as cancer.
• Examples of diseases that are diagnosed using
FISH include Prader-Willi syndrome, Angelman
syndrome, chronic myelogenous leukemia,
acute lymphoblastic leukemia, Cri-du-chat,
Velocardiofacial syndrome, and
Down syndrome
 DNA/RNA
ANALYSIS

DNA EXTRACTION
DNA PURIFICATION
POLYMERASE CHAIN
DNA AMPLIFICATION REACTION
DNA CLEAN-UP
DNA ANALYSIS

DUCHENNE MUSCLE DYSTROPHY

• Thank you