Laboratory diagnosis cancer

1. LABORATORY DIAGNOSIS OF
CANCER
Dr Utkarsh Sharma

2. Common techniques of cancer
diagnosis
1. Radiological diagnosis
2. Cytological diagnosis
3. Histological diagnosis
4. Frozen section
5. Haematogical diagnosis
6. Immunohistochemistry
7. Molecular diagnosis
8. Tumour markers

3. 1.RADIOLOGICAL DIAGNOSIS
It includes
• X-ray
• Ultrasound
• CT scan
• MRI
• These are one of the best non invasive
techniques for early diagnosis of cancer.

6. 2.CYTOLOGICAL DIAGNOSIS

7. • 1. Fine needle aspiration cytology (FNAC)
• Fine needle aspiration cytology is a popular
method of tumor diagnosis particularly for
palpable tumors
• Lymph nodal tumors
• Breast tumors
• Salivary gland tumors
• Thyroid tumors
• Liver SOL

12. 3.HISTOLOGICAL DIAGNOSIS :
• For histological diagnosis the following methods
of sampling is done:
• Biopsy- biopsy is a surgical removal of small
piece of tissue for microscopic examination for
the presence of cancer cell.
There are three ways tissues can be removed for
• Biopsy:-
• Endoscopy
• Needle biopsy
• Surgical biopsy

13. • Endoscopy- in this process , a thin, flexible tube with a
tiny camera on the end is inserted into the body cavity.
This allows the doctor to view the abnormal area.
• Needle biopsy - the doctortakes a small tissue sample
by inserting a needle into abnormal area. Different
types of needles are used, Ex:
• Vim Silverman needle for liver biopsy.
• Renal biopsy needle for renal tissue.
• True cut biopsy needle for prostatic tissue or breast
tissue.
Surgical Biopsy:-
• There are two types of surgical biopsies.
• An excisional biopsy: it is performed when the doctor
removes the
entire tumor, often with some surrounding normal tissue.

14. • An incisional biopsy: it is performed when the doctor
removes just a portion of the tumor. If cancer is found to
be present, the entire tumor may be removed immediately
or during another operation. The processing of tissue and
its diagnosis takes a two or three days.

15. 4. FROZEN SECTION:-
Frozen section is quick diagnosis method. The tissue
is quickly frozen at around -20 C in frozen section
cryostat which makes the tissue hard.
-tissue is immediately sectioned & stained
-the whole process from receiving, staining to diagnosis
can be completed within 10 to 15 days.

17. 5. HAEMATOLOGICAL DIAGNOSIS:-
• Complete blood count (CBC)
• Peripheral blood film (PBF)
• Bone marrow aspiration
• Bone marrow imprint smear
• Bone marrow biopsy.
Important in diagnosis of leukemia.

18. Flow cytometry
 Flow cytometry is a technique used to detect and
measure physical and chemical characteristics of a
population of cells or particles.
 A sample containing cells or particles is suspended in
a fluid and injected into the flow cytometer instrument.
 The sample is focused to ideally flow one cell at a time
through a laser beam and the light scattered is
characteristic to the cells and their components.
 Cells are often labeled with fluorescent markers so that
light is first absorbed and then emitted in a band of
wavelengths. Tens of thousands of cells can be quickly
examined and the data gathered are processed by a
computer.

20. 6.

22. 7.MOLECULER DIAGNOSIS
Molecular diagnostics is a collection of techniques
used to analyse biological markers in the genome and
proteome—the individual's genetic code and how their
cells express their genes as proteins by applying
molecular biology to medical testing.

23. Allele-Specific PCR
Variants detected: Typically considered targeted
analysis for the detection of a specific single nucleotide
variant (SNV).
Time to complete test: 1–2 days.
Pros: Sensitive—can detect mutant DNA if present at
1–5%. No special equipment required.
Cons: Target specific and cannot detect
other mutations that may be present in tumor DNA.

24. Sanger Dideoxy Sequencing
Variants detected: Unknown mutations including
SNVs and small duplications, insertions, deletions, and
indels of interest.
Pros: A variety of unknown mutations can be detected.
Can be used to detect gene fusions if RNA from the fusion
transcript is first extracted from the specimen. No special
equipment required.
Cons: Labor intensive and requires mutant DNA to be
present at 20–25%. Cannot detect changes in exon
or gene copy number.

25. Pyrosequencing
Variants detected: Unknown mutations in a small
targeted region.
Time to complete test: 2–3 days.
Pros: Quick and sensitive detection of mutant DNA at a
level of 5%.
Cons: Requires pyrosequencing instrumentation; is
limited in the types of mutations that can be detected in
tumor DNA.

26. Mass Spectrometry – MS
Variants detected: Targeted SNVs.
Time to complete test: 2–3 days.
Pros: Sensitive, reliably detecting mutant DNA if
present at 5–10%; tests more than one gene.
Cons: Requires mass spectrometry instrumentation;
SNV-specific and cannot detect other mutations in
tumor DNA that may be present.

27. Multiplex Ligation-Dependent
Probe Amplification – MLPA
Variants detected: Exon and gene copy
number. Depending on experimental design, can
also detect SNVs.
Pros: Quick and able to detect
multiple mutations simultaneously. Can detect targeted
SNVs at 10%.
Cons: For exon or gene copy number variant detection,
requires mutant DNA to be present at levels of 20–40%.
Testing works better on fresh frozen tissue than
on DNA extracted from paraffin-embedded tissue.

28. Fluorescence In Situ Hybridization
Variants detected: Targeted gene copy number changes
and targeted SVs.
Method: Fluorescent probes are used to locate genes or
sequences of interest on one or more chromosome.
Fluorescence microscopy is used for detection.
Time to complete test: 2–3 days.
Pros: Easily detects gene copy number changes and
targeted SVs that are not as easily detected by other
methods; cell-based imaging enables detection of events
in a small fraction of cells.
Cons: Requires paraffin-embedded tissue on unstained
slides; cannot detect most types of mutations occurring in
solid tumor neoplasms.

29. Next Generation
Sequencing- NGS
A sequencing method where millions of sequencing
reactions are carried out in parallel, increasing the
sequencing throughput.
Pros: Enables simultaneous detection of single base
substitutions as well as more
complex mutations including duplications, insertions,
deletions, and indels in many genes in a single assay;
requires low inputs of DNA.
Cons: Expensive

31. 8. TUMOR MARKER:-
• Some tumors release certain substances called
tumor markers.
• Blood test can be performed to detect the blood cells
as well as for specific tumormarkers.
• Tumor marker is biochemical indicators
of tumors, these may be:
• Antigens
• Cytoplasmic proteins
• Enzymes
• Hormones
Used in support diagnosis

32. Liquid biopsy
Tumors almost always shed some amount of their
fragments into peritumoral space.
These fragments may be represented by single malignant
cells or their clusters as well as by various proteins,
nucleic acids, small molecules, etc.
Consequently, these entities can be collected in various
body fluids (serum, saliva, urine, etc.) and serve as
tumor markers.