Maj Gen (R) Suhaib Ahmed  HI (M)Maj Gen (R) Suhaib Ahmed, HI (M)MBBS; MCPS; FCPS; PhD (London)

Genetics Resource Centre (GRC) Genetics Resource Centre (GRC) Rawalpindi

www.grcpk.com

Sickle Cell Anemia: a Molecular Disease.P li  & I li  S i  N b  Pauling & Italiano. Science November 1949

Watson & Crick: 1953

l h i i ( )Polymerase Chain Reaction (PCR)

(Kary Mullis 1985)

DNADNA

chromosome

cell nucleus

Target Region for PCRTarget Region for PCRDouble stranded DNA molecule

IndividualIndividual nucleotides

Molecular Pathology: ApplicationsInherited DisordersCancerInfectious DiseasesHuman IdentificationOthers

Of the 5000 known Genetic Disorders2500 can be diagnosed by Genetic Analysis

P t l Di iIdentify Couple at Risk

Prenatal DiagnosisIdentify Couple at Risk

Pregnancy

Fetal Sampling

Lab DiagnosisAffected Fetus Normal Fetus

Termination of Pregnancy

Ch i i Vill S li (CVS)Chorionic Villus Sampling (CVS)

Prenatal Diagnosis of Thalassaemia

700

800

500

600

200

300

400

0

100

200

0

1994

1995

1996

1997

1998

1999

2000

2001

2002

2003

2004

2005

2006

2007

2008

2009

2010

2011

2012

2013

Molecular Diagnosis of CancerMolecular Diagnosis of Cancer

AFIP

Molecular MarkersMolecular Markers• CML bcr-ablCML bcr abl• ALL bcr-abl, Ig-gene, TCR-gene • AML M2 t(8;21)AML M2 t(8;21)• AML M3 t(15;17)• AML M4Eo inv(16)• AML M4Eo inv(16)• NHL bcl-II, Ig-gene, TCR-gene• MPD JAK2 mutation• MPD JAK2 mutation

JAK2 Mutation

Real Time PCR for Bcr‐abl

50%

100%

25%

12.5%

6.25%

3.12%

Negative

1.6%<1%

Infectious DiseasesViral

HCV, HBV, HIV, CMV, HPV, Dengue etc.Bacterial

TuberculosisA tibi ti   i tAntibiotic resistance

ParasiticLeishmania  MalariaLeishmania, Malaria

Others

R Q

5’3’

3’

5’

Primer-F 3’5’ Taqman Probe(A)

5’ 3’

3’ Primer-R5’

R Q

5’3’

3’Primer-F 3’

5’

(B)

5’ 3’

3’ Primer-R5’

Q

5’3’

3’Primer-F

R(C)

5’

5’

3’ Primer-R5’

3’

Real Time PCR for HCVReal Time PCR for HCV

Malaria

Human Identification(Forensic DNA Testing)

Donor Chimerism

DLI

A Mobile DNA LabA Mobile DNA Lab

GTI-16 DNA Thermal Cyclery

GTI 16 ABI 9700 (USA)GTI-16 ABI-9700 (USA)

GTI-PCR Reader-GGTI-PCR Reader-G

GTI-Microcentrifuge & Dry Bath

GTI-DC Power Supply

GTI PAGE A tGTI-PAGE Apparatus