Chromosomes and Karyotyping

Dr. Nisrine Bissar-Tadmouri

Chromosomes
Karyotype: display of chromosomes paired according to their size, location of the centromere, and staining patterns. organized into 7 groups

Cytogenetics a discipline that matches phenotypes with detectable chromosomal abnormalities.

Male karyotype

Trisomy 21 or Downs Syndrome

Submetacentric
Metacentric

Acrocentric

Diagnosis of chromosome abnormalities Child born take blood and look at lymphocytes Unborn child Prenatal Diagnosis Chorionic villus sampling (CVS) Amniocentesis (AF) Fetal Blood Sampling (FBS) 1) Karyotype 2) FISH: Fluorescent in situ Hybridization

Indications for Karyotyping

An individual with characteristics of one of the common chromosomal syndromes (Downs, Turners, kleinfelter etc..) The presence of multiple congenital anomalies (usually associated with mental retardation) A couple with multiple miscarriages An individual at risk for inheriting or transmitting a chromosomal rearrangement (translocation) A child with ambiguous external genitalia

Downs syndrome FISH is used for fine details like submicroscopic deletions, or even gene level mutations (hard to detect on the Karyotype)

Screening Both in first and second trimester Maternal serum screening Ultrasound Indicates which should go forward for invasive procedure

Human Karyotype preparation for prenatal diagnosis

Around 16 weeks

10-12 weeks

How to prepare a chromosome spread

sample 0.7 ml heparinized blood 10ml cell growth medium containing phytohemagglutinin (mitogen) Incubate 72 hrs at 370C

synchronization 100 ml colcemid Incubate for 20 min at 370C

Cell collection Centrifuge at 1200 rpm for 10 min Remove medium leaving 0.5 ml

swelling Resuspend cells in the remaining medium Add 2 ml of 0.075M KCl drop by drop Add 8 ml of the KCl solution Incubate for 15 min at 370C

fixation Centrifuge and remove supernatant Resuspend cells and add 10 ml freshly prepared fixative (methanol: glacial acetic acid, 3:1) for 10-15 min Centrifuge and remove supernatant

Metaphase spread Resuspend in small amount of fixative Drop suspension on microscop slide Stain by immersing in Giemsa stain 7-10 min

Writing karyotype information

Abbreviation
46, XY 46, XX 45, X 47, XXY 47, XYY 46, XY del (7q) 47, XX+21

What it means
Normal male Normal female Turner syndrome female Klinefelter syndrome male Jacobs syndrome male Male missing part of long arm of chromosome 7 Female with trisomy 21

46, XY t (7;9) (p21.1;q34.1)

Male with translocation between short arm of chromosome 7 band 21.1 and long arm of chromosome 9 band 34.1

Writing karyotype information

Abbreviation
46, XX, del(5)(p15.3) 46, XY, dup(21)(q21) 45, XX, inv(9)(p12q13) 46, XX,del(15)q12 46, XX/45,X0

What it means
Cri du Chat syndrome female Male with duplication of band q21 on chr 21 Female with percentric inversion on chr 9 Female with Prader-Willi A mosaic case of Turner Syndrome