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Imse Lab > ANA DETECTION > Flashcards

ANA DETECTION Flashcards

(56 cards)

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1
Q

Materials

A

⤳ SLE latex suspension
⤳ Black test cards
⤳ Mechanical rotator
⤳ Positive and Negative Controls
⤳ Disposable stirrers

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2
Q

Principle of the test

A

Passive Latex Agglutination

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3
Q

Meaning of ELISA

A

Enzyme-Linked Immunosorbent Assay

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4
Q

Meaning of CLIA

A

Chemiluminescence immunoassay

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5
Q

These assays can testfrom a broad range of antibodies if multiple nuclear antigens are coated onto a single test well, or for a specific
ANAs if each well is coated with a single antigen.

A

ELISA and CLIA

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6
Q

ELISA & CLIA depends on?

A

Depends on the type of nuclear antigen loaded on the well

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7
Q

Capable of detecting various diseases

A

Multiple nuclear antigens

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8
Q

Capable of detecting single or specific diseases

A

Specific nuclear antigen

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9
Q

These antigens use in commercial kits are derived from

A

Tissue extracts or produced by recombinant technology

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10
Q

It is used to screen for the presence of anas in addition to identifying specific anas

A

ELISA methods

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11
Q

There is a large variation in the performance of test produced by different manufacturers, which is influenced by the

A

Antigen preparation used

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12
Q

One study found sensitivities of elisa assays ranging from

A

69% to 98%

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13
Q

Specificities of elisa ranges from

A

81% to 98% when they were compared with IFF ANA method

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14
Q

Proportion of patients with a disease who test positive

A

Sensitivity

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15
Q

Proportion of patients without a disease who test negative

A

Specificity

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16
Q

If 100 patients known to have a disease were tested, and 43 test positive, then the test has 43% sensitivity. If 100 with no disease are tested and 96 return a completely negative result, then the test has 96% specificity.

A

True

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17
Q

Reagents

A

Ana reagent
Positive and negative control reagents
Wells

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18
Q

Qualitative method (screening) procedure

A
  1. Bring all reagents to room temperature and mix gently prior to use.
  2. Place the following on separate divisions of the same black test card.
  3. Place a drop of undiluted serum into the circles of the test slides
    ⤳ Patient’s Serum: 1 drop
    ⤳ Positive Control: 1 drop
    ⤳ Negative Control: 1 drop
  4. Add 1 drop of SLE Latex reagent to each sample on the black test card.
  5. Mix with the flat end of the pipette/mixer and spread fluid evenly covering the entire
    diameter of the test circle.
  6. Tilt the slide back and forth slowly for 3 minutes while observing for agglutination.
  7. Observe and Interpret results.
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19
Q

The test is considered as NEGATIVE when no difference in agglutination is observed between the specimen and negative control and POSITIVE sera must show distinct agglutination within _ minutes.

A

3

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20
Q

Agglutination indicates the level of ___ (especially anti-DNP) in the
range commonly found in the SLE.

A

antinuclear antibody

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21
Q

Findings in the sensitivity studies suggest that immunoassays may miss a significant proportion of ANA-positive patients and also yield a significant number of _____

A

false positive results.

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22
Q

Based on such studies, the ___ has recommended that the IIF test remains the gold standard for ANA testing and that clinical laboratories should specify the method they use when they are reporting the result.

A

ACR (American College of Rheumatology)

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23
Q

gold standard for ANA testing and that clinical laboratories should specify
the method they use when they are reporting the result.

Study These Flashcards
A

Indirect Immunofluorescence Test
(IIF)

24
Q

FANA TESTING PATTERNS

Study These Flashcards
A

HOMOGENOUS
SPECKLED
NUCLEOLAR
MIXED
CENTROMERE
PERIPHERAL

25
CLINICAL SIGNIFICANCE
Screening method for SLE but not specific
26
is a chronic systemic inflammatory disease that affects people usually at the peak age between 20 and 40 year
Systemic Lupus Erythematosus
27
SLE is a chronic systemic inflammatory disease that affects people usually at the peak age between
20 and 40 years
28
Women are much likely more to be affected than men by a ratio of ___.
9:1
29
This autoimmune disease appears to originate from complex interaction between several factors, such as:
Environmental Genetic susceptibility Abnormalities within the immune system.
30
⤳ In line with these, over ___ autoantibodies associated with SLE have been discovered.
100
31
Autoantibodies includes
● double stranded DNA (dsDNA) ● histones, and other nuclear components, as well as ● autoantibodies to lymphocytes, erythrocytes, platelets, phospholipids, ribosomal components, and endothelium
32
Laboratory diagnosis of SLE include
CBC, platelet count and urinalysis.
33
Findings in lupus patients are
leukopenia and possible anemia and thrombocytopenia.
34
Erythrocyte Sedimentation Rate (ESR) may be ___ even though C-Reactive Protein tends to be low or normal.
Elevated
35
When SLE is suspected, the first test typically done is a screening test for Antinuclear Antibodies (ANA). These antibodies are present in most cases of SLE, antibodies that are directed against antigens of
nuclei of mammalian cells.
36
ANAs are present in over _____ of patients with active SLE and are used as a major marker for the disease.
95%
37
ANAs are not specific only to SLE as it may be present in other connective tissue diseases such as
Sjogren’s syndrome Scleroderma Polymyositis-dermatomyositis RA.
38
They can also be found in some individuals with other conditions, including:
Chronic infections Cancer Pregnancy
39
Furthermore, up to ___ of healthy individuals and up to ___ of elderly people are ANA-positive
5% 30%
40
are a heterogeneous group of antibodies that have different antigen specificities.
ANAs
41
The nuclear antigens they are directed against
- double stranded DNA - single stranded DNA - histones - nucleosomes - centromere proteins - extractable nuclear antigens
42
These antibodies are the most specific for SLE because they are mainly seen in patients with lupus and their levels correlate with disease activity.
Double stranded DNA (DsDNA)
43
Antibodies to dsDNA typically produces what type of staining pattern on indirect immunofluorescence (IIF)?
peripheral or homogenous
44
Methods in detecting ANAs have been developed. ⤳ These include the ff:
- Indirect Immunoflurescence test (IIF) - Immunoperoxidase staining - Enzyme linked immunosorbent assay (ELISA) - Microsphere multiplex immunoassays (MIA) - Radioimmunoassay(RIA) - Immunodiffusion - Immunoblotting (Western blot) - Dot blot - Immunoelectrophoresis - Microarray.
45
OTHER IMPORTANT INFORMATION ● Note the proper procedure and usage of the __
reagents, storage temperature and the expiry date of the kit.
46
Pattern in fana
Peripheral (rim) Homogeneous (diffused) Speckled Centromere Nucleolar
47
Antibodies associated with peripheral pattern
Anti-DNA (not seen in HEp-2)
48
Disease associated with peripheral pattern
SLE
49
Antibodies associated with homogeneous pattern
Anti-DNA Anti-histone Anti-DNP (Dinitrophenyl)(nucleosomes)
50
Disease associated with homogeneous pattern
Rheumatoid Arthritis Systemic Lupus Erythymatosus Miscellaneous Disorders (anti-ssDNA)
51
Antibodies associated with speckled pattern
Anti-sm and RNP Anti- Ro & La Anti-Jo-1 and Mi-2 Anti- Scl-70
52
Diseases associated with speckled pattern
SLE and Systemic Sclerosis (SS) Polymyositis (PS)/ Dermatomyositis (DM) PSS (Progressive Systemic Scleroderma)
53
Antibody associated with centromere pattern
Anti-centromere
54
Disease associated with centromere pattern
Progressive systemic scleroderma CREST - Calcinosis - Raynaud's phenomenon - Esophageal dysfunction - Sclerodactyly - Telengiectaisas
55
Antibody in nucleolar pattern
Anti- nucleolar
56
Diseases associated in nucleolar pattern
SLE & PSS (Progressive Systemic Sclerosis)

Imse Lab (2 decks)

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