NGS for diagnosis of Epilepsy

Syndromes

Dr. Pragasam Viswanathan

Professor, SBST
 Epilepsy can be defined as a chronic seizure disorder or group
of disorders characterized by seizures that usually recur
unpredictably in the absence of a consistent provoking factor.

Epilepsy is not contagious

it is not a mental illness
or a cognitive disability.

The neurological dysfunction seen in epilepsy can :

 begin at birth
 childhood

 adolescence or

 even in adulthood
DIAGNOSIS
• Electroencephalogram
(EEG)
• Neurological imaging studies
*Magnetic Resonance Imaging
(MRI)
*Functional MRI (fMRI)
*Computed Tomography (CT)
*Positron emission tomography
(PET)
*Single-photon emission
computerized tomography
(SPECT)
 DNA SEQUENCING
 DNA Sequencing is Figuring out the order of
DNA nucleotides, or bases (A T G C ), in a genome
that make up an organism’s DNA.
Sangar
Sequencing

F. Sangar
 ERA OF SEQUENCING
1st Generation sequencing ABI PRISM 377
• Sequence many identical molecules
• Sequencing in large gels or capillary tubing
limits scale

Sangar Chain Termination Maxam- Gilbert Sequencing

( 1977 ) (1977)
 ERA OF SEQUENCING
5

1st Generation Sequencing 2nd Generation Sequencing

• Sequence many identical • Sequence millions of

molecules clonally amplified molecules
per run QIAGEN GeneReader
• Sequencing in large gels or
capillary tubing limits scale • Using a reversible,
stepwise sequencing
chemistry
• Immobilized on a surface

Illumina MiSeq
Life Technologies/Applied Roche / 454
Biosystems; SOLID 5500 Pyrosequencer
Intro to NGS, 11.30.2016
NEXT GENERATION SEQUENCING
High throughput DNA Sequencing Technique.
Employs Micro and Nanotechnologies
Reduce sample size.
Low Reagent cost
Less Time
Massive Parallel Sequencing
Sequence thousands of sequences at once.
Produce enormous amount of data .
 NGS WORKFLOW
Sample Extraction , DNA fragmentation and invitro adapter ligation

Clonal Amplification by Clonal Amplification by

Emulsion PCR Bridge PCR

Pyrosequencing Sequencing-by-ligation Sequencing-by-synthesis

(454 Sequencing) (SOLiD Platform ) ( Solexa Technology)