Transcript Molecular diagnostics

Molecular Diagnostics at
The Hospital for Sick Children
Molecular Diagnostic Laboratory
•12 Technologists
• 30 tests
• diagnosis, carrier testing
Molecular Diagnostic Laboratory
Achrondroplasia
Angelman Syndrome
Apert Syndrome
Batten Disease (NCL)
Beckwith-Wiedemann syndrome
Canavan disease
Craniosynostosis, non-syndromic
Crouzon/Jackson-Weiss syndrome
Cystic Fibrosis
Duchenne/Becker Muscular Dystrophy
Fabry disease
Familial Dysautonomia
Fragile X syndrome
FRAXE
Gaucher disease
Hearing loss, non syndromic
Hunters disease
HHT
Hypochondroplasia
Pfeiffer syndrome
Prader-Willi syndrome
Retinoblastoma
Saethre-Chotzen syndrome
Schwachman-Diamond Syndrome
Spinal and bulbar muscular atrophy
Spinal Muscular Atrophy
Tay-Sachs disease
Thanatophoric Dysplasia (Type I & II)
Bone Marrow Transplant Testing
DNA extraction/banking
Identity testing
X-inactivation
Y-specific marker (ZFX/ZFY)
Zygosity Testing
Types of Mutations Tested
Point mutations?
Few recurrent
mutations?
Many unique
mutations?
Disease
Other
mutations?
Deletions &
duplications?
Whole gene?
Some exons?
Also with point
mutations?
Diagnosis of Batten Disease
Neuronal Ceroid Lipofuscinoses (NCL)
Primer Extension Test for NCL Mutations
Normal
Mutant
C451T
Multiplexed Analysis of 9 NCL Mutations
C70G
IVS
C622T
1.02del
Normal
delAT
C451T A364T
A223C
G284V
Duchenne Muscular Dystrophy (DMD)
DMD Mutation Types
60
%
40
20
Deletion Duplication Point
Deletions in DMD Gene
BMD
Intermediate
DMD
3'
5'
1
10
20
30
40
50
60
70
Exons
DMD Deletion Carrier Analysis
DMD Deletion/Duplication Analysis – 79 exons
exon
DMD A
DMD F
5, 15, 25, 32,
43, 52, 63, 73
2, 16, 34, 42,
53, 64, 78
DMD B
DMD G
DMD C
DMD H
7, 14, 23, 38,
46, 54, 62
BP, 6, 17, 26,
33, 45, 55, 65,
75
10, 18, 27, 35,
44, 56, 66, 76
3, 13, 22, 37,
47, 58, 67, 74
DMD D
DMD I
1, 11, 21, 30,
40, 50, 60, 71
9, 19, 28, 36,
48, 57, 68, 77
DMD E
4, 12, 24, 31,
41, 51, 61, 72
exon
DMD J
8, 20, 29, 39,
49, 59, 69, 79
DNA Sequencing
Normal
Carrier
Automated Mutation Detection
ABI SeqScape