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Molecular Testing in Cancer

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Anil Tarigopula
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15 views

Molecular Testing in Cancer

Uploaded by

Anil Tarigopula
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
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Molecular Testing in

Cancer
The Why & How
Dr Anil Tarigopula MD, DNB
Senior Consultant,
Apollo Hospitals, Chennai
Is Molecular testing “ESOTERIC”?

• Laboratory medicine - the most analytical of the


medical specialties
• Strong focus on technology and result reporting to
provide data for clinical decision making
• The type of lab testing must be taken in the context of
patient outcomes and not as a sole reflection of the
aspects of the test system
Molecular Testing in Clinical
Diagnostics
Analysis of nucleic acids from tissue, cells or fluids to
diagnose disease
predict the occurrence of disease
predict the prognosis of diagnosed disease
guide therapy
Molecular Techniques - capable of detecting DNA/RNA
in very small quantities in tissue and body fluids,
enabling detection of various disease states
Accuracy and as short TAT as possible
Practicality of testing and clinical needs need
consideration
Important to use standardized methods and apply
established standards of Quality Assurance
Results MUST NOT be used alone but in coordination
with traditional laboratory tests
Molecular Diagnostics-advantages
• Address limitations of conventional diagnostics by taking
advantage of technological advances
• Emerging molecular diagnostic assays-
• Rapid turnaround times
• Minimal hands on time
• High sensitivity & specificity
• Significantly higher positive rates than by conventional methods
• High negative predictive value

• Eliminate guesswork in Clinical Practice


Molecular Diagnostics- The Process
of testing
Sample collection and processing
Sample types
Sample collection/storage
DNA extraction methods
RNA extraction methods
Nucleic acid storage / handling
Amplification
Signal amplification
Target-based amplification
Signal detection methods
DNA binding dyes
Probes
Non-amplification methods
Hybrid capture
In situ hybridization
Allele specific & sequence specific oligonucleotide hybridization
Reverse hybridization
Molecular Diagnostics- The Process of
testing
• DNA separation methods
• Agarose gel electrophoresis
• Polyacrylamide gel electrophoresis
• Capillary electrophoresis
• Sequencing of nucleic acids
• Sanger sequencing
• Pyrosequencing
• Next-gen sequencing

Necessary to understand:
1. Target
2. Matrix
3. Method
4. Assay format
Molecular Testing Process
• Requesting molecular analysis
• Preanalytical standards
• Sample receipt and handling
• DNA and RNA extraction
• Choice of analytical method
• Performance of molecular tests
• Reporting molecular diagnostic tests for patients
• QA – internal and external
• Training and expertise
• Research and development
Cree IA, et al. J Clin Pathol 2014; 0:1-9
Polymerase chain reaction

Biochemical technology in molecular biology used


to amplify a single copy or a few copies of a piece
of DNA across several orders of magnitude, generating
Molecular Lab – Test Portfolios

• Molecular Infectious Disease

• Molecular Oncology

• Transplantation diagnostics

• Molecular Pathology of Inherited disease

• Pharmacogenomics
SAMPLE QUALITY
Molecular Oncology
FISH

Mutation status

Molecular profiling of disease

Disease Load monitoring


NGS based testing in
Oncology
Dr Anil Tarigopula MD, DNB
Consultant
Molecular Diagnostics and Transplantation Immunology
Apollo Hospitals, Chennai
Clinical Testing
• Personalised medicine - Accurate, reproducible and
clinically useful diagnostic tests to identify patients
who can benefit from targeted therapies

• Test (s) - must provide information that is essential


for the safe and effective use of a corresponding drug
or biological product

• Test Validation:
• Analytical validation
• Clinical validation
NGS based Oncology
Diagnostics
• Most of the current CDx- single analyte / pre-specified
mutations
• NGS based testing - numerous biomarkers concurrently
• Simultaneous detection of
• Single nucleotide variants
• Copy number alteration
• Insertions
• Deletions
• Translocations
• Drug repurposing - Application of known/approved drugs to
new indications/cancer type - New target discovery
• Ability to process pooled multiple samples and perform
parallel sequencing, allowing for an appreciable cost
reduction.
PROCESSING
From Specimen To Result: Many Areas of Pre-Analytic Variability
From Specimen To Result: Many Areas of Pre-Analytic Variability
Quality Assurance Program

Environment Personnel Equipment Supplies Specimens Internal External Accurate and


- Safe and - Trained and - Maintained - Uninterrupted - Good quality quality Quality timely
functional competent and serviced supplies Labelled with monitoring Assessment reporting
- Temperature staff - Appropriate unique ID - Test working (EQA) - Turnaround
control - Test user is transport and - Completed properly - Testing site‘s time
documented storage request form work checked Results review
- Current SOPs conditions by another
readily site
available

Accurate, reliable and timely results


THANK YOU

29

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