Genetics-Index

SL Topics List Previous Year Page

No.
1 Chromosome R-14
2 Nucleic acid, Gene, Genetic code, Karyotyping
3 Mutation
4 Genetic Disease
5 Mendelian Disorder
• Autosomal Dominant Disorders R-20, 17
R-20, 19, 18, 16, 15, 14
• Autosomal Recessive Disorders D-20, 16, 12
R-18, 17, 15, 14, 12
• X-linked Recessive Disorders R-17, 15, 11 & D-20, 19, 15
6 Multifactorial inheritance & Disorders D-19
7 Cytogenetic Disorders R-19, 18, 17, 16, 15 & D-19
i) Down Syndrome R-17, 13, 12, 09
ii) Edward’s Syndrome
iii) Patau’s Syndrome
iv) Turner’s Syndrome R-12, 10
v) Klinefelter’s Syndrome R-19, 18, 13, 12 & D-20, 18
8 Non-Disjunction
9 Chromosome Analysis
10 Diagnosis of genetic diseases R-20, 14 & D-17
11 Molecular Methods R-17, 15 & D-20
 Chromosome

Number of chromosome is constant:

 Each human somatic cell is 46 number of chromosome (Diploid or 2n)

 Number of chromosome in a ova or sperm (23 or n haploid)
 44 autosome, 2 sex chromosome

Size:

✓ Visible when cell is in mitotic or meiotic cell division

✓ Average size of chromosome in metaphase is 5-micron meter

Shape: V or rod J shaped

Structure: Short arm is ‘p’, Long arm is ‘q’.

Centromere:

 Responsible for movement of chromosome during cell division, towards specific and
essential region
 Visible after condensation
 Consist of DNA and proteins, spindle fibers are attached
 In anaphase centromere divides longitudinally into two sister chromatids and moves to
opposite poles.

Nucleic acid, Gene, Genetic code, Barr body

• Nucleic acid is responsible for inheritance

• DNA is found in nucleus (chromosome) and mitochondria

• Components of DNA:
Deoxyribose sugar + phosphoric acid + nitrogenous bases (A, T, C, G)

• Purines → A, G
• Pyrimidines → C, T

Basic terms:

• Nitrogenous base + Sugar = Nucleoside

• Nucleoside + Phosphate = Nucleotide

• Polymer of nucleotides = Nucleic acid

Components of Nucleic Acid:

A) Sugar

i) Deoxyribose → only in DNA

ii) Ribose → only in RNA

B) Nitrogenous Bases

1. Purines

• Adenine (DNA & RNA)

• Guanine (DNA & RNA)

2. Pyrimidines

• Cytosine (DNA & RNA)

• Thymine (only DNA)
• Uracil (only RNA)

RNA

90% is present in cytoplasm and 10% in nucleolus

Three forms:

1. mRNA is the template for polypeptide synthesis. It has a cap at 5′ end, and a poly A tail
at 3′ end.
2. tRNA brings activated amino acids into position along mRNA template.
3. rRNA is a component of ribosomes which functions as a non-specific site of polypeptide
synthesis.

Gene

Gene is the basic unit of inheritance for a given characteristics. Gene is the shortest segment of
chromosome within which genetic information is stored.

A typical gene includes:

1. A promoter region- binds RNA polymerase for the initiation of transcription

2. A transcription initiation site
 3. A translation initiation site- Designate the first AA in the protein
4. A transcription termination codon
5. 3′ untranslated region
o Stabilize mRNA
o Allows it to exit nucleus
o Permits to be translated

Different between DNA and RNA:

DNA RNA
DNA found in nucleus (chromosome) mitochondria RNA found in nucleolus, ribosome
and cytoplasm
Sugar is deoxyribose Sugar is ribose
Double stranded Single stranded
Four nitrogenous base (A–adenine, T-thymine, C- A,U (uracil) C,G
cytocine, G–guanine)
Hereditary material Non hereditary, helps in protein
synthesis

Genetic code

 Genetic code directs transcription of RNA and translation into protein.

 Codons are read in 5′ → 3′ direction.
 A codon = 3 nucleotides coding for one amino acid.
Example: ATG → Methionine.
 Total 64 triplet codons:
o 61 code for amino acids
o 3 are stop (nonsense) codons → TAA, TAG, TGA

Characteristics:

1. Specificity → One codon codes for one specific amino acid.

2. Universality → Same genetic code in almost all organisms.
3. Redundancy (Degeneracy) → Multiple codons may code for same amino acid.
4. Non-overlapping & Commaless → Read continuously, 3 bases at a time without
punctuation.

Barr Body

 Barr body (sex chromatin) = inactive X chromosome seen in female interphase nuclei.
 It is genetically inactive heterochromatin.

Characteristics:

 Seen during interphase, disappears during cell division.

  Found in cheek epithelial cells & neutrophils (drumstick appearance).

Number of Barr Body:

Barr body = Number of X chromosomes − 1

Indications for Barr Body Count:

a. Ambiguous genitalia
b. Females with:

 Lymphedema in newborn
 Primary amenorrhoea
 Inguinal mass
 Features of Turner syndrome
 X-linked disorders

c. Males with:

 Mild–moderate mental retardation

 Severe hypospadias
 Cryptorchidism
 Small firm testes
 Infertility
 Gynaecomastia

Barr body examination helps diagnose:

 46, XX
 47, XXY (Klinefelter syndrome)
 49, XXXXX
 49, XXXXY

Karyotyping

Definition: Study of chromosomal constitution of a cell/individual

Normal male karyotype = 46, XY

Steps of Karyotyping:

1. Isolation of nucleated cells (blood lymphocytes/fibroblasts)

2. Cell culture
3. Metaphase arrest by colchicine
4. Separation of dividing cells
 5. Fixation with methanol + glacial acetic acid
6. Micro-photography

Uses of Karyotyping:

 Detect chromosomal abnormalities

 Detect numerical abnormalities
 Detect structural abnormalities

Mutation

Definition: Permanent change in DNA sequence

involving coding region of DNA.

 Mutations may cause disease or may be

harmless.
 Harmless sequence variations are called
genetic polymorphism.

Causes:

1. Errors in replication
o Mispairing of bases
o Insertion of extra nucleotide
2. Errors in recombination / rearrangement
3. Irradiation
o X-ray, γ-ray, UV ray
4. Exposure to mutagens
o Base analogues
o Alkylating agents
o Nitrous oxide
o Free radicals
5. Spontaneous alteration or loss of bases
o Example: Deamination of
cytosine → uracil

Types of Mutations:

1. Genomic Mutation : Gain or loss of whole chromosome

o Numerical abnormality
2. Chromosomal Mutation : Structural chromosome changes
o Example: deletion, insertion, translocation
3. Gene Mutation
A. Point Mutation: Single base change

Types:

 Silent mutation → Same amino acid formed despite codon change

 Missense mutation → Different amino acid formed
o Example: Sickle cell anemia
 Nonsense mutation → Premature stop codon
o Example: β-thalassemia

B. Frame Shift Mutation

 Insertion/deletion of 1 or 2 bases
 Not multiple of 3
 Alters reading frame
 Example: Duchenne muscular dystrophy

C. Deletion / Insertion of 3 or Multiples of 3: Abnormal protein synthesis without

frame shift

D. Triplet Repeat Mutation: Expansion of repeated nucleotide sequences

Features:

 May occur in normal individuals

 Premutation carriers may be asymptomatic
 Expansion may increase in next generation (anticipation)

Examples

Disease Type
Huntington disease Coding repeat expansion
Myotonic dystrophy Non-coding expansion
Fragile X syndrome Non-coding expansion
Friedreich ataxia Non-coding expansion

Genetic disease
Classification of Genetic Diseases:

Category Description
Mendelian disorders Due to mutation in a single gene with large effect; follows
classic Mendelian inheritance
Multifactorial disorders Caused by interaction of genetic + environmental factors;
involves multiple genes with small additive effects
Cytogenetic disorders / Due to chromosomal abnormalities
Chromosomal abnormalities

Types of Chromosomal Mutations / Abnormalities:

Type Subtype Examples

Genomic/ Numerical Aneuploidy → Gain or loss of Monosomy, Trisomy,
mutation one or more chromosomes Tetrasomy
Monosomy → Loss of 1 Turner syndrome
homologous chromosome
Trisomy → Gain of 1 Trisomy 21, 18, 13
homologous chromosome
Tetrasomy → Gain of 2 —
homologous chromosomes
Polyploidy → Addition of Triploidy, Tetraploidy
complete haploid chromosome
sets
Structural / Translocation Burkitt lymphoma → t(8;14),
Chromosomal Philadelphia chromosome →
mutation t(9;22)
 Deletion → Loss of nucleotide Wilms tumor
segment
Insertion → Addition of —
nucleotide segment
Inversion Paracentric, Pericentric
Ring chromosome —
Isochromosome —

Chromosomal & Contiguous Gene Disorders:

Disorder Karyotype Key Features

Down syndrome 47,XX/XY,+21 Intellectual disability, congenital heart disease
(Trisomy 21)
Edwards syndrome 47,XX/XY,+18 Early death, skull/facial abnormalities, organ
(Trisomy 18) defects
Patau syndrome 47,XX/XY,+13 Cleft lip/palate, polydactyly, CHD, early
(Trisomy 13) lethality
Klinefelter syndrome 47,XXY Male infertility, gynecomastia, small testes
XYY syndrome 47,XYY Usually asymptomatic, behavioral issues
Triple X syndrome 47,XXX Usually asymptomatic, mild ↓ IQ
Turner syndrome 45,X Short stature, webbed neck, primary
amenorrhea, coarctation of aorta

Microdeletion / Contiguous Gene Syndromes

Syndrome Locus Key Features

DiGeorge / 22q11.2 Cardiac defects, cleft palate, hypocalcemia
Velocardiofacial
Prader–Willi syndrome 15q11–q13 Hyperphagia, obesity, small hands/feet
Angelman syndrome 15q11–q13 Ataxia, absent speech, abnormal EEG
Williams syndrome 7q11.23 Supravalvular AS, learning disability,
hypercalcemia
Smith–Magenis syndrome 17p11.2 Behavioral abnormalities, sleep disturbance
 Mendelian disorder(classical single gene disorder)

 There is gene mutation. These disorders are due to defect in a single gene.
 Mode of inheritance follows Mendelian principles.
 Mutant gene may be located in autosomes or in sex chromosomes.

Four Pattern of Inheritance:

 Autosomal dominant
 Autosomal recessive
 X-linked dominant
 X-linked recessive

Autosomal Dominant vs Autosomal Recessive:

Feature Autosomal Dominant (AD) Autosomal Recessive (AR)

Genotype Usually heterozygous Usually homozygous
Anticipation May occur Usually absent
Parents At least one parent affected Parents usually carriers
Carrier state Absent Present
Sex distribution Male & female equally Male & female equally affected
affected
Transmission Vertical transmission Horizontal transmission
pattern
Risk to offspring 50% offspring affected 25% affected if both parents are
carriers
Generation pattern No generation gap Generation gap present
Onset & prognosis Late onset, better prognosis Early onset, poorer prognosis
Defect commonly in Structural proteins / receptors Enzyme proteins
Mutation New mutation may occur Often associated with consanguinity
Expression Variable expressivity Usually complete penetrance

Autosomal Dominant Disorders

Characteristics:

 Usually manifest in heterozygous state

 Both sexes equally affected & transmit disease
 Affected × unaffected parent → 50% risk in each child
 Usually one affected parent present
 May occur due to new mutation
 Commonly involve structural protein defects
 Show reduced penetrance & variable expression
 Often late onset (e.g., Huntington disease)
  No carrier state
 Vertical transmission (no generation skipping)

Examples of Autosomal Dominant Disorders:

System Disorders
Nervous Huntington disease, Neurofibromatosis, Myotonic dystrophy, Tuberous
sclerosis
Urinary Polycystic kidney disease
Gastrointestinal Familial polyposis coli
Hematopoietic Hereditary spherocytosis, Von Willebrand disease
Skeletal Marfan syndrome, Ehlers-Danlos syndrome, Osteogenesis imperfecta,
Achondroplasia
Metabolic Familial hypercholesterolemia, Acute intermittent porphyria
Others Long QT syndrome, Brugada syndrome, Hereditary hemorrhagic
telangiectasia, Charcot-Marie-Tooth disease

Autosomal Recessive Disorders

Characteristics:

 Manifest only when both alleles are mutated (homozygous state)

 Males & females equally affected
 Parents are usually asymptomatic carriers
 Horizontal transmission (often seen in one generation)
 Each pregnancy carries 25% recurrence risk
 Common in consanguineous marriage
 Usually due to enzyme defects
 Often show complete penetrance
 Typically early onset with severe disease
 Many involve inborn errors of metabolism
 Incidence ≈ 25/1000 live births

Examples of Autosomal Recessive Disorders:

System Disorders
Metabolic Cystic fibrosis, Phenylketonuria, Galactosemia, Albinism, Lysosomal
storage diseases, Wilson disease, Hereditary hemochromatosis
Hematopoietic Sickle cell anemia, β-thalassemia
Endocrine Congenital adrenal hyperplasia
Skeletal Alport syndrome, Ehlers–Danlos syndrome (some variants), Ataxia
telangiectasia, Muscular atrophies
Nervous Friedreich ataxia, Neuronal ceroid lipofuscinosis, Peroxisomal disorders
Others Kartagener syndrome, Alkaptonuria, Mediterranean fever
 Sex-Linked Disorders

 Mutant gene is located on the sex chromosome.

 All sex-linked disorders are usually X-linked.
 Most are X-linked recessive.
 Y chromosome is relatively resistant to mutation.

X-linked Recessive Disorders

Characteristics:

 Females are usually carriers and males are usually affected.

 Affected males receive the mutant X chromosome from the mother.
 Y chromosome has no normal allele to modify the effect.
 Affected males transmit the mutant gene to all daughters but not to sons.
 Females are usually carriers.

Female may be affected if:

 Homozygous mutant female

(affected father + carrier mother)
 Turner syndrome
 Lyon hypothesis (skewed X inactivation)
 Affected male may have normal parents
(carrier mother + healthy father)
 Disease commonly runs in maternal lineage
(maternal uncle → nephew)

Examples of X-linked Recessive Disorders:

System Disease
Musculoskeletal Duchenne muscular dystrophy
Blood Hemophilia A & B
Chronic granulomatous disease
G6PD deficiency
Immune Agammaglobulinemia
Wiskott–Aldrich syndrome
Metabolic Diabetes insipidus
Lesch–Nyhan syndrome
Nervous Fragile X syndrome

X-Linked Dominant Disorder

Characteristics:
  Affect both sexes but female > male
 No male-to-male transmission
 Affected father transmits disease to all daughters but not to sons
 No carrier state

Examples of X-Linked Dominant Disorders:

 Vitamin D resistant rickets (Hypophosphatemic rickets)

 Rett syndrome
 Coffin–Lowry syndrome
 Oral-facial-digital syndrome type I
 Incontinentia pigmenti
 Periventricular hypertrophy
 Xg blood group
 Alport syndrome

Mode of Transmission:

Inheritance Key Features

Autosomal dominant Structural gene defects; many generations affected; both sexes
affected
Autosomal recessive Usually enzyme deficiencies; carrier state present; often seen in one
generation
X-linked recessive Sons of carrier mothers have 50% risk; no male-to-male
transmission; skips generations
X-linked dominant Mother transmits to 50% of sons & daughters; affected father → all
daughters affected, no sons
Mitochondrial Transmitted only through mother; all offspring of affected female
inheritance may be affected

Comparison between X-linked Dominant & X-linked Recessive:

Feature X-linked Dominant X-linked Recessive

1. Affected father × normal All daughters affected, no Daughters become carriers,
mother sons affected no sons affected
2. Carrier / affected mother × 50% sons & 50% daughters 50% sons affected, 50%
normal father affected daughters carriers
3. Transmission pattern Vertical transmission; no Skipped generation may
generation gap occur
4. Sex affected Females more affected than Mostly males affected
males
5. Severity Often fatal in males Females usually spared or
mild
6. Male-to-male transmission Absent Absent
 Multifactorial inheritance & disorder

Multifactorial inheritance is more common than monogenic inheritance.

It results from interaction of multiple genes + environmental factors → also called polygenic
inheritance.

Features of Multifactorial Inheritance:

 Expression depends on number of genes involved

 ~5% risk in 1st-degree relatives
 Concordance in identical twins is >5% but far <100%
 Risk increases if more siblings are affected
 Can involve:
o Continuous traits → height
o Discontinuous traits → diabetes

Common Multifactorial (Polygenic) Disorders:

 Cleft lip ± cleft palate

 Congenital hip dislocation
 Congenital heart disease
 Hypertension
 Gout
 Diabetes mellitus
 Neural tube defects (anencephaly, spina bifida)
 Pyloric stenosis
 Talipes (club foot)
 Exomphalos (omphalocele)

Examples of Multifactorial Disorders:

 Disorder
 Asthma
 Schizophrenia
 Congenital heart disease
 Epilepsy (idiopathic)
 Mental retardation (idiopathic)
 Spina bifida
 Congenital pyloric stenosis
 Anencephaly
 Type-1 diabetes mellitus
 Club foot
 Cleft lip ± palate
Sibling Risk (λs) in Common Polygenic Diseases:

Disease λs
Multiple sclerosis 20–40
Type 1 diabetes mellitus 15
Schizophrenia 10
SLE 10–20
Ischaemic heart disease 4–12

Cytogenic/Chromosomal disorder

These disorders may occur due to abnormalities in:

1. Number of chromosomes
2. Structure of chromosomes

Numerical Chromosomal Disorders:

1. Polyploidy: Chromosome number is an exact multiple of the haploid number

 Examples: 69, XX, 92, XX

2. Aneuploidy: Chromosome number is increased or decreased

 Not an exact multiple of haploid set

 Example: 47, XXX

Aneuploidy(Gain or loss of one chromosome)

 Monosomy → Absence of one chromosome (2n−1)

 Trisomy → Presence of an extra chromosome (2n+1)
 Mosaicism → Presence of two cell populations; some normal and others with
missing/extra chromosome

Examples of Aneuploidy:

Autosomal Sex Chromosomal

Trisomy: Trisomy:
Down syndrome Klinefelter syndrome
Patau syndrome Super male (XYY)
 Edward syndrome Super female (XXX)
Monosomy: Monosomy:
Usually not compatible with life Turner syndrome

Structural Chromosomal Abnormalities:

Type Description
Translocation Transfer of chromosomal segment to another chromosome; may be
reciprocal or Robertsonian
Deletion Loss of chromosomal segment
Insertion Addition of chromosomal segment
Isochromosome Centromere divides transversely instead of longitudinally
Inversion Two breaks followed by inversion and reinsertion
Ring Fusion of damaged chromosome ends after terminal deletion
chromosome

Trisomy 21 or Down’s syndrome

Karyotype:

 47,XX/XY,+21
 Translocation forms: 21/22, 21/21

Incidence & Risk:

 Most common chromosomal disorder

 Incidence ≈ 1 in 800 births
 Risk increases with maternal age
o ~1:1500 (<20 years)
o ~1:25 (>45 years)

Causes:

Cause Frequency Notes

Nondisjunction ~94–95% Sporadic; maternal age related
Robertsonian translocation ~4% Due to parental chromosomal abnormality
Mosaicism ~1–2% Milder clinical features

Clinical Features of Down Syndrome:

General

 Intellectual disability
  Developmental delay
 Short stature
 Hypotonia

Facial Features

 Flat face & occiput

 Epicanthic folds
 Upward slanting eyes
 Brushfield spots
 Low-set ears
 Macroglossia
 Short neck

Musculoskeletal

 Single palmar crease

 Short broad hands
 Clinodactyly
 Sandal gap
 Joint laxity
 Atlanto-axial instability

Cardiac Defects

 AVSD (most common)

 ASD, VSD, PDA
 Tetralogy of Fallot

GIT Abnormalities

 Duodenal atresia
 Hirschsprung disease
 Umbilical hernia

Associated Disorders

 Congenital hypothyroidism
 Leukemia (ALL/AML)
 Pulmonary hypertension

Screening for Down Syndrome:

Trimester Screening Test / Marker Result

 First trimester (11–14 weeks) Nuchal translucency ↑ Increased
β-hCG ↑ Increased
PAPP-A ↓ Decreased
Second trimester (13–20 weeks) (Triple hCG ↑ Increased
test)
α-fetoprotein (AFP) ↓ Decreased
Unconjugated estriol (uE3) ↓ Decreased

Edward Syndrome (Trisomy 18)

Clinical Features:

 Intellectual disability
 Prominent occiput
 Low-set ears
 Micrognathia
 Rocker-bottom feet
 Congenital heart disease
 Finger deformities with overlapping fingers
 Death usually within first few days/months

Mnemonic: EDWARDS

 E → Elongated occiput
 D → Digits overlapping
 W → Wide head
 A → Absent intellect
 R → Rocker-bottom feet
 D → Diseased heart
 S → Small lower jaw

Patau Syndrome (Trisomy 13)

Clinical Features

 Intellectual disability
 Cleft lip & palate
 Hare lip
 Polydactyly
 Microphthalmia
 Congenital heart disease

Comparison: Trisomy 13 vs Trisomy 18:

Feature Trisomy 13 (Patau) Trisomy 18 (Edward)

Head & Cleft lip/palate, microphthalmia, Prominent occiput, micrognathia,
 Face holoprosencephaly low-set ears
Extremities Polydactyly Overlapping fingers, rocker-bottom
feet
Chest CHD common CHD common
General Severe developmental delay Severe developmental delay
Prognosis Poor survival Poor survival

Turner Syndrome

Definition: Complete or partial monosomy of X chromosome

 Phenotypic female with hypogonadism

 Most common sex chromosome abnormality in females

Karyotype:

 45,X (most common)

 Barr body absent
 May show mosaicism or structural X abnormality

Clinical Features:

System Features
General Short stature, delayed puberty,
primary amenorrhea, infertility,
failure of secondary sexual development
Genital / Gonadal Infantile genitalia, poor breast development,
sparse pubic hair, streak ovaries / ovarian atrophy
Face & Neck Low-set ears, high-arched palate,
webbed neck, low posterior hairline
Chest & Skeletal Shield chest, widely spaced nipples,
increased carrying angle, short 4th metacarpal/metatarsal
Cardiovascular Coarctation of aorta, bicuspid aortic valve,
aortic stenosis, hypertension
Renal Horseshoe kidney,
collecting system anomalies
Neurologic Usually normal intelligence,
mild visuospatial/nonverbal defects
Others Lymphedema (non-pitting edema), hypothyroidism,
hearing loss, pigmented nevi, ↓ bone density,
impaired glucose tolerance
 Klinefelter Syndrome

Definition: Male with one or more extra X chromosomes

Most common cause of hypogonadism in males

Karyotype:

 47,XXY (most common)

 One Barr body present
 Usually due to meiotic nondisjunction
 Mosaic type: 46XY/47XXY

Clinical Features:

System Features
General Eunuchoid body habitus, long legs
Secondary Sexual Sparse facial/body hair, high-pitched voice, poor beard growth
Features
Genital Small penis, small atrophic testes, infertility
Endocrine Hypogonadism, gynecomastia, ↑ breast cancer risk
Cognitive Intelligence average to mildly reduced
Metabolic Type 2 diabetes, metabolic syndrome, insulin resistance
Musculoskeletal Osteoporosis
Cardiovascular Mitral valve prolapse, ASD/VSD
Others ↑ risk of extragonadal germ cell tumors, autoimmune disease
(e.g., SLE)

Investigations

 ↑ FSH, LH & estrogen

 ↓ Testosterone
 Azoospermia

Non-Dysjunction

Definition: Failure of homologous chromosomes to separate during meiosis

Causes of Non-Disjunction:

Advanced Maternal Age

 Maternal age >35 years ↑ risk of trisomy 21

  Primary oocyte remains arrested for years before meiosis II

Other Causes

 Radiation
 Delayed fertilization after ovulation
 Smoking, alcohol
 OCPs, fertility drugs, pesticides
 Genetic factors

Maternal Age Also Affects:

 Hydrocephalus
 Anencephaly
 Achondroplasia (also related to paternal age)

Chromosome Analysis

Prenatal Chromosome Analysis:

Indications

 Maternal age >34 years

 Previous child with chromosomal abnormality
 Determine fetal sex in X-linked disorders

Postnatal Chromosome Analysis:

Indications

 Multiple congenital anomalies

 Suspected aneuploidy (e.g., Down syndrome)
 Intellectual disability / developmental delay
 Suspected chromosomal abnormality

Diagnosis of Genetic Diseases

Prenatal Diagnosis

Usually performed in families with:

 Previous affected child

 Advanced maternal age
 Positive family history
  Abnormal screening test

Methods:

Method Details
Fetal cells Obtained from amniotic fluid or chorionic villi for
chromosomal/enzyme analysis
Biochemical analysis Estimation of AFP, enzyme levels, metabolites in amniotic
fluid
Chorionic Villus Sampling Done at 8–10 weeks; biopsy from chorionic villi for
(CVS) chromosomal/DNA study

Diseases Diagnosed Prenatally:

System Diseases
Musculoskeletal Becker muscular dystrophy,
Duchenne muscular dystrophy, Myotonic dystrophy
Neurologic Tay–Sachs disease, Neurofibromatosis, Neural tube defects
Metabolic G6PD deficiency, Phenylketonuria
Chromosomal Down syndrome, Turner syndrome, Klinefelter syndrome
Hematologic β-thalassemia, Sickle cell anemia
Others Huntington disease, Cystic fibrosis

Indications for Prenatal Diagnosis:

 Maternal age >35 years

 Previous affected child
 Positive family history
 Abnormal maternal screening tests
 Parents carrying genetic/chromosomal disorders
 X-linked disorders with male fetus

Maternal Screening Markers in Down Syndrome

Marker Change
AFP ↓
uE3 (unconjugated estriol) ↓
hCG ↑
PAPP-A ↓

Methods of Prenatal Diagnosis:

Method Gestational Age Main Use

 Ultrasound 1st trimester Detects fetal anomalies & nuchal
onward translucency
Chorionic Villus 9–12 weeks Early chromosomal/DNA analysis;
Sampling (CVS) miscarriage risk ~2%
Amniocentesis 14–16 weeks Chromosomal & biochemical analysis;
miscarriage risk <1%
Cordocentesis 18–20 weeks Fetal blood/DNA analysis; miscarriage risk
2–3%

Postnatal Diagnosis

Usually performed on blood lymphocytes.

Methods:

Method Details
Karyotyping Study of complete chromosome set; metaphase arrest with colchicine
Barr body Inactive X chromosome seen in female cells; used for sex chromatin analysis

Molecular Methods

Molecular Methods / Techniques:

Major Categories

1. Enzymatic digestion
2. Hybridization
3. Gel electrophoresis
4. Amplification methods

Amplification Methods:

 Cloning (genetic recombination)

 PCR
 NASBA (nucleic acid sequence–based amplification)
 LCR (ligase chain reaction)
 TMA (transcription-mediated amplification)

Hybridization

Based on binding of complementary nucleic acid strands to form a hybrid double strand.
Blotting Techniques

Blot Detects
Southern blot DNA
Northern blot RNA
Western blot Protein

Polymerase Chain Reaction (PCR)

Technique for amplification of specific DNA sequences.

Types

Type Function
RT-PCR Amplifies RNA after conversion to complementary DNA (cDNA)
Real-time PCR Detects & quantifies nucleic acid

Applications of PCR:

1. Prenatal diagnosis (e.g., cystic fibrosis)

2. Diagnosis of hereditary diseases (sickle cell anemia, hemophilia)
3. Diagnosis of infections (viral, bacterial, fungal, parasitic)