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ANA Test

The patterns seen are: 1. Homogeneous: Possible SLE or drug-induced lupus 2. Nucleolar: Possible systemic sclerosis 3. Speckled: Possible SLE, Sjögren's syndrome or MCTD

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Sami Al Wesabi
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117 views16 pages

ANA Test

The patterns seen are: 1. Homogeneous: Possible SLE or drug-induced lupus 2. Nucleolar: Possible systemic sclerosis 3. Speckled: Possible SLE, Sjögren's syndrome or MCTD

Uploaded by

Sami Al Wesabi
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Clinical immunology

(Lab. 2)

ANA test
Dr. Sami Al-wesabi
Anti-Nuclear Antibodies (ANA)
 Antinuclear antibodies (ANAs, also known as
antinuclear factor or ANF) are autoantibodies that bind
to contents of the cell nucleus.

 ANAs are present in over 95% of patients with active


lupus and are used as a major marker for the disease.
They can also be found in 5% of healthy persons and up
to 30% of elderly individuals are ANA-positive.
Anti-Nuclear Antibodies (ANA)
This allows the use of ANAs in the diagnosis of some autoimmune
disorders, including :
 Systemic Lupus Erythematosus(SLE).

 Sjögren's Syndrome(SS).

 Scleroderma/Systemic Sclerosis .

 Mixed Connective Tissue Disease(MCTD).

 Polymyositis(PM).

 CREST syndrome ( Calcinosis, Raynaud’s syndrome, Esophageal syndrome,


Telangiectasia)

 Autoimmune Hepatitis(AIH).

 Drug induced lupus.


Anti-Nuclear Antibodies (ANA)
Subtypes of ANAs:
There are many subtypes of ANAs such as:
1. Anti-Double-stranded DNA antibodies(Anti-dsDNA ).

2. Anti-histone antibodies.

3. Anti-centromere antibodies.

4. Anti-nuclear ribonucleoprotein (anti-nRNP)

5. Anti-Smith (Anti-Sm) antibodies. Extractable nuclear antigens (ENA):


They are termed ENA because they can be extracted
6. Anti-Scl-70 antibodies. from the cell nucleus with saline. The ENAs consist of
ribonucleoproteins and non-histone proteins, named by
7. Anti-Jo-1 antibodies. either the name of the donor who provided the
prototype serum or the name of the disease setting in
8. Anti-Ro/SS-A antibodies. which the antibodies were found.

9. Anti-La/SS-B antibodies.
Anti-Nuclear Antibodies (ANA)
Methods used for ANA detection:
 ELISA technique.
 Immunofluorescence technique.

 ANA ELISA testing.

Principle:

Enzyme-linked immunosorbent assay uses antigen-coated micro titter


plates for the detection of ANAs. Each well of a micro titter plate is
coated with either a single antigen or multiple antigens to detect specific
antibodies or to screen for ANAs
Anti-Nuclear Antibodies (ANA)
ANA ELISA testing.

 Interpretation of ANA result:

ANA-ELISA results is read as positive or negative.


Anti-Nuclear Antibodies (ANA)
 The immunofluorescence ANA test
 Indirect immunofluorescence is one of the most commonly used tests for
ANAs.

Principle:

HEp-2 cells are used as a substrate to detect the antibodies in human


serum. Microscope slides are coated with HEp-2 cells and the serum is
incubated with the cells. If antibodies are present then they will bind to
the antigens on the cells nucleus. These can be visualized by adding a
fluorescent tagged (usually (FITC) ) anti-human antibody that binds to
the antibodies. The molecule will fluoresce when a specific wavelength
of light shines on it, which can be seen under the microscope.
Anti-Nuclear Antibodies (ANA)
 The immunofluorescence ANA test

 Interpretation of ANA pattern:

ANA-IFA results are reported as a titter and a fluorescence pattern. Depending on the
antibody present in the human serum and the localization of the antigen in the cell
Anti-Nuclear Antibodies (ANA)
Autoantibody Antigen (s) Pattern Disease Association
Anti-dsDNA dsDNA Homogeneous SLE

Anti-histone Histone Homogeneous Drug-induced SLE


Anti-Nuclear Antibodies (ANA)
Autoantibody Antigen (s) Pattern Disease Association
Anti-Sm Extractable Speckled SLE
nuclear antigen
(RNA component)

Anti-RNP Proteins Speckled 1- SLE


complexed with
nuclear RNA 2- MCTD
Anti-Nuclear Antibodies (ANA)
Autoantibody Antigen (s) Pattern Disease Association
Anti–Scl-70 DNA Nucleolar Systemic sclerosis.
topoisomerase I

Anti–Jo-1 Histidyl-tRNA Fine cytoplasmic speckling Polymyositis


synthetase.
Anti-Nuclear Antibodies (ANA)
Autoantibody Antigen (s) Pattern Disease Association
Anti-SS-A (Ro) Proteins speckled 1- SLE.
complexed to
RNA 2-Sjögren’s syndrome.

Anti-SS-B (La Phosphoprotein Finely speckled 1- SLE.


complexed to
RNA polymerase 2-Sjögren’s syndrome.
Anti-Nuclear Antibodies (ANA)
Autoantibody Antigen (s) Pattern Disease Association

Anti-centromere Centromere Centromere CREST syndrome.


Anti-Nuclear Antibodies (ANA)
B C
A
Anti-Nuclear Antibodies (ANA)
F G
E
Q/ from the following clinical pictures, diagnose the pattern
and possible disease?

1 2 3

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