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Chapter 5 Principles of Clinical Cytogenetics

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33 views32 pages

Chapter 5 Principles of Clinical Cytogenetics

Uploaded by

r28327047
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Principles of Clinical

Cytogenetics and Genome


Analysis
Chapter 5
INTRODUCTION TO CYTOGENETICS AND GENOME
ANALYSIS

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INTRODUCTION TO CYTOGENETICS AND
GENOME ANALYSIS
• Chromosome disorders form a major category of
genetic disease.
• Reproductive wastage
• Congenital malformations
• Intellectual disability
• Play an important role in the pathogenesis of cancer

• Cytogenetic abnormalities in 1% of live births


• 2% of pregnancies in women older than 35 years
• Fully half of all spontaneous, first-trimester
abortions
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Sample sources
• Fibroblasts from skin biopsy
• White blood cells – lymphoblastoid
• Bone marrow – a high proportion of proliferative cells
- invasive procedure - hematological malignancies
• Fetal cells derived from amniotic fluid (amniocytes) or
obtained by chorionic villus biopsy can also be
cultured successfully for cytogenetic, genomic,
biochemical, or molecular analysis.
• Fetal cells –
• amniotic fluid (amniocytes)
• chorionic villus biopsy
• Cultured successfully for cytogenetic, genomic,
biochemical, or molecular analysis.
• Chorionic villus cells can also be analyzed directly
after biopsy, without the need for culturing
• Cell-free fetal
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4
plasma
CLINICAL INDICATIONS FOR CHROMOSOME AND
GENOME ANALYSIS

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Chromosome
Identification

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Chromosome Identification
• Giemsa banding (G banding)
• Metacentric chromosomes – more or less central
centromere
• Submetacentric chromosomes, off-center centromere,
and arms of clearly different lengths
• Acrocentric chromosomes, with the centromere near
one end.
• Telocentric ??
• Acrocentric chromosomes (chromosomes 13, 14, 15,
21, and 22)
• Satellites
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and narrow stalks (called secondary
7
constrictions) –rRNA
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The Karyotype: an international description
Total number of chromosomes,

Sex chromosome constitution,

Anomalies/variants.
46,XY
47,XX,+21 Trisomy 21 (Down syndrome)
47,XXX Triple X syndrome
69,XXY Triploidy

45,XX,der(13;14)(p11;q11) Robertsonian translocation


46,XY,t(2;4)(p12;q12) Reciprocal translocation

46,XX,del(5)(p25) Deletion tip of chromosome 5


46,XX,dup(2)(p13p22) Duplication of part of short arm Chr 2
46,XY,inv(11)(p15q14) Pericentric inversion chromosome 11
46,XY,fra(X)(q27.3) Fragile X syndrome
46,XY/47,XXY Mosaicism normal/Klinefelter syndrome
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Chromosome Identification
• High-resolution
banding –
(prometaphase
banding)
• 850 bands or even
more in a haploid
set.
• Diagnosis of
microdeletion
syndromes
• 2- to 3-Mb size
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range
Fluorescence In Situ
Hybridization
• Detecting - a
particular DNA
sequence
• evaluating the number
or organization of a
chromosome or
chromosomal region in
situ
• Molecular
cytogenetics,
cytogenomics, or
chromonomics
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Genome Analysis Using
Microarrays
• Comparative genome
hybridization (CGH)
Microarrays
• https://youtu.be/MMkYSWzOwrY
• https://youtu.be/qmefXRNFsq8
• https://youtu.be/bRML0PkH-wE
• https://youtu.be/0URm6Ta6v2k

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Microarrays
uses
Chromosome and genome
abnormalities in
children with
unexplained
developmental delay
intellectual disability
birth defects
revealing several
pathogenic genomic
alterations that are not
detectable by (G
banding) karyotyping

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Genome Analysis by Whole-
Genome Sequencing
• Sequence patient genomes in their entirety.
• Aneuploidies
• Chromosomal rearrangements
• Birth defects
• Cancer
• Variations (SNPs)
• https://youtu.be/rH6vsoyrc_U
• https://youtu.be/P0xUb-rRxU4

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CHROMOSOME ABNORMALITIES

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Gene Dosage, Balance and
Imbalance
• Chromosome and genomic disorders-quantitative
aspects of gene expression
• Single-gene disorders-qualitative aspects of a
gene’s function
• Gene dosage and its balance or imbalance.

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Abnormalities of Chromosome
Number
• Heteroploid = other than 46
• Euploid = diploid (2n)
• Aneuploid 2n+1…OR -1…
• Triploidy (3n) – (partial molar pregnancy)
• 1% to 3% of recognized conceptions
• dispermy or diploid egg or sperm
• Partial hydatidiform mole
• Tetraploidy (4n)
• 92,XXXX or 92,XXYY
• Failure of completion of an early cleavage division
of the zygote.
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Aneuploidy
• Trisomy 21 - Down syndrome (karyotype 47,XX,+21
or 47,XY,+21)
• Trisomy 18-
• Trisomy 13-
• Monosomy - Turner syndrome
• Meiotic nondisjunction.
• Mosaicism

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Detection of trisomy 21 in a
female patient by whole-genome
chromosomal microarray

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Detection of trisomy 21 by whole-
genome sequencing and
overrepresentation of sequences from
chromosome 21.

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Abnormalities of Chromosome
Structure
Unbalanced Rearrangements
• Duplication of part of a chromosome leads to
partial trisomy for the genes within that
segment;
• Deletion leads to partial monosomy
• Detected by
• High-resolution karyotyping (chromosomal banding)
• FISH
• Chromosomal microarray analysis

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Abnormalities of Chromosome
Structure
Unbalanced Rearrangements
• Deletions – partial monosomy
• Terminal
• Interstitial
• Duplications - partial trisomy
• Marker (supernumerary chromosomes) and Ring
Chromosomes
• Isochromosomes
• Dicentric Chromosomes
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Abnormalities of Chromosome
Structure
Balanced Rearrangements
• Translocations
• Reciprocal Translocations
• Robertsonian Translocations rob(13;14)(q10;q10)
• Insertions
• Inversions
• Paracentric
• Pericentric

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Mosaicism for Chromosome
Abnormalities
• The cause of
mosaicism is
nondisjunction in an
early postzygotic
mitotic division.
• Germline Mosaicism
Occurs when all or part
of a parent’s germline
is affected by a
disease-causing
mutation

• Pseudo-mosaicism
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Incidence of Chromosome
Anomalies
• Spontaneous
Abortions- 40% to
50%
• 45,X - Turner
syndrome - nearly
20% of chromosomally
abnormal spontaneous
abortuses
• 1% of chromosomally
abnormal live births

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