The

role of Cytogenetics in the medical laboratory

BHSc in Medical Laboratory Science Cytogene5cs CYT200A Deni5on: Cytogene5cs is the analysis of blood or bone marrow cells that reveals the organiza5on of chromosomes. Chromosomes are the physical structures that contain the gene5c material, DNA. Samples brought to the cytogene5cs laboratory are studied to aid in the diagnosis of inherited condi5ons, and to diagnose or monitor cancer condi5ons with associated chromosome abnormali5es.

The Time Line: The dark ages prior to 1952 2n=48 The hypotonic era 1952 2n = 46 The trisomy period Lejeune describes Down syndrome extra chr 21 The Banding era dieren5al staining The Molecular era Higher resolu5on screening Need to understand the Cell Cycle -For culture --to understand gene5c abnormali5es caused during cell division

Mitotic inhibitor arrests the spindle formation and stops cell division at metaphase: Chromatin is the protein-DNA complex in which genetic material exists in the interphase nucleus. During metaphase, chromatin condenses into chromosomes, which can be stained and analysed.

Normal Male Karyotype

Block stained metaphase and karyotype

Cytogenetics Laboratory

Prenatal Samples Increased Trisomy Risk

(triple test) Abnormal Ultrasound Family history of chromosome abnormality

Blood Samples
Recurrent Miscarriages Infertility Family History of Chromosome abnormalities Ambiguous Genitalia Learning difficulties and delayed development Dysmorphic Features

Solid Tissue Samples Recurrent miscarriages

Congenital abnormalities

Oncology Samples
Haematological and other solid tumour cancers: Bone marrow, blood samples, tumour tissue, tumour effusion

Forensic Samples
Varied, for molecular analysis

Karyotyping

100 Mbp

10 Mbp

1 Mbp

100 Kbp

10 Kbp

1 Kbp

100 bp

10 bp

1 bp

Size of Abnormality

Array CGH FISH Karyotyping MLPA

100 Mbp

10 Mbp

1 Mbp

100 Kbp

10 Kbp

1 Kbp

100 bp

10 bp

1 bp

Size of Abnormality

Spermatogonium

Meiosis
Primary spermatocyte

Oogonium

Primary oocyte

Genetic variation through crossover and recombination

Secondary spermatocytes

Secondary oocyte

Polar Body I 4 spermatids

Polar Body II

4 spermatozoa

Fetal period
Mitosis

Oogonium

Females

Before or at birth
Meiosis in progress

After birth
Arrested in diplotene of Meiosis I

Primary Oocyte

After puberty
Meiosis I complete Arrest at Metaphase II

Secondary Oocyte & Polar Body I

At fertilization
Meiosis II complete

Fertilized Ovum & Polar body II

Dierences in Gametogenesis
Male
Puberty 60-65 days 30-500 mitoses 4 sperma5ds 100-200 million /

Female
Early embryonic development 10-50 years 20-30 1 ovum and polar bodies 1 ovum / menstrual cycle

ejaculate

Parental origin of aneuploidy

Paternal % 15 10 5 80 5 45 Maternal % 85 90 95 20 95 55 0 Trisomy 13 Trisomy 18 Trisomy 21 45,X 47,XXX 47,XXY 47,XYY

100

Down syndrome
95% standard trisomy 1% mosaics Due to increase in maternal age 4% transloca5ons no age eect

Chromosome abnormalities in miscarriages

Trisomy 13 Incidence% Incidence%

2 15 3 5

Other Trisomy 25 Monosomy X 20 Triploidy Tetraploidy Other 15 5 10

Trisomy 16 Trisomy 18 Trisomy 21

Chromosome abnormalities in newborns

Incidence / 10,000 births
Trisomy 13 Trisomy 18 Trisomy 21 45,X 47,XXX 47,XXY 47,XYY Unbalanced Balanced Total 2 3 15 1 10 10 10 10 30 90 Patau SYndrome Edwards Syndrome Down syndrome Turner Syndrome Triple X Syndrome Klinefelter Syndrome

Chromosome Abnormalities
Triploidy ---- rare at birth lethal Trisomy 16 --- most common in spontaneous miscarriages,

completely lethal.

Trisomy 13 & 18 95% miscarry Trisomy 21 80% miscarry Klinefelters 50% miscarry 45X 1% at concep5on. 98 % miscarry, mosaic survives

Non-disjunction during meiosis

18

Down Syndrome karyotype

19

46,XY,+21 (describes a pure trisomy

ISCN Nomenclature
p arm Centromere q arm

20

q10

1. Number 2. Sex 3. Abnormality

1. Identification of a chromosome by length and position of the centromere. 2. Accurate reporting of abnormalities (GPS co ordinates) 3. The smaller the defect, the longer the chromosome needs to be for detection

Cytogenetics: Application of ISCN Trisomy Information to Karyotyping: Down Syndrome Patau Syndrome Edwards Syndrome Other than the established culture and karyotype of chromosomes we will also cover some molecular techniques: FISH Fluorescent in situ hybridisation PCR polymerase chain reaction