FIANLS - ANA - DETECTIO OF ANTI-NUCLEAR ANTIBODIES Flashcards
materials used for ana
sle latex suspension
positive and negative control
black test cards
mechanical rotator
disposable stirrers
procedure
add 1 drop of SLE latex reagent to each sample on the black test card
mix the flat end of the pipette or mixer and spread the fluid evenly covering the entire diameter of the test circle.
tilt the slide back and forth slowly for 3 minutes while observing for agglutination
observe and interpret the results
principle of the test
ELISA and CLIA - automated pocess
ELISA and CLIA are assays that can test from a broad range of antibodies if multiple nuclear antigens are coated onto a single test well, or for specific ANAs if each well is coated with a single antigen. These antigens used in commercial kits are derived from tissue extracts or produced by recombinant technology. Because of their advantages, man laboratories are using ELISA methods to screen for presence of ANAs in addition to identifying specific ANAs
ELISA means
enzyme-linked immunosorbent assay
CLIA means
Chemiluminescence Immunoassay
is sensitivity of ELISA depends on the machine and the manufacturer?
yes
one study found that the sensitivity of ELISA assay is ranging from __ to ___ and specificity from ___ to ___ when they were compared with IIF ana method
sensitivity- 69% to 98%
specific - 81% - 98%
what do we look for if we’re conducting a qualitative test/
agglutination
do we need to bring all the reagents first to room temp before using
yes, then mix gently prior to use
it indicates a positive result
agglutination
the positive sera must show distinct agglutination within __mns
3 mns
agglutination indicates the level of ___in the range commonly found in the SLE
antinuclear antibody (especially anti dnp)
findings in the sensitivity studies suggest that immunoassays miss a significant proportion of ____ and also yield a significant number of false positive results.___
ANA-positive patients; false positive results
based on such studies, the ACR has recommended that the ___remains the gold standard for ANA testing and that clinical laboratories should specify the method they use when they are reporting the result.
IIF test - indirect immunofluorescent test
is a chronic systemic inflammatory disease that affects people usually at the peak age between 20 - 40
systemic lupus erythematosus
is ANA a confirmatory test?
nope
ANA can be detected in multiple auto immune disease, so it’s not ___
specific
SLE is at peak at ages ___
20 - 40
what gender is most likely to be affected by SLE
women
ratio of how women are most likely to be affected by SLE than men
9:1
this auto immune disease appears to originate from complex interaction between several factors, such as ___
environmental, genetic susceptibility, and abnormalities within the immune system
over ___ autoantibodies associated with SLE have been discovered
100
among the 100 auto antibodies found in SLE,
it include
double stranded DNA (ds DNA), histones, and other nuclear components,
among the 100 sle antibodies, there are auto antibodies to formed elements such as
erythrocytes, and platelets, as well as other body components such as phospholipids, ribosomal components and endothelium
laboratory diagnosis of SLE include ___
CBC, platelet count and urinalysis
findings in Lupus patients are ___
leukopenia, and possible anemia, and thrombocytopenia.
esr in SLE may be ___ even though C reactive protein tends to be low or normal
elevated
the most specific antibodie for SLE
double stranded DNA
when SLE is suspected, the first test typically done is a screening test for __
anti-nuclear antibodies
these antibodies are present in most cases of SLE, antibodies that are directed against antigens of nuclei of mammalian cells
true or false
true
ana’s are present in over __ of patients with active SLE and is used as a major marker for the disease
95%
ANAs are not specific only to SLE as it may be present in other connective tissue disease such as ___, ___, _, and ___
Sjogren’s syndrome, scleroderma, polymyositis-dermatomyositis, and RA
ANAs, they can also be found in some individuals with other conditions including ___
chronic infections, pregnancy, ad cancer.
up to ___of healthy individuals is positive to ana
5%
up to ___of elderly people are positive to ANA
30%
ANAs are ____ group of antibodies that have different antigen specificities
(heterogenous, homogenous)
heterogenous
The nuclear antigens they are directed against include ____
double or single stranded DNA, histones nucleosomes, centromere proteins, ad extractable nuclear antigens
why do double stranded dna antibodies are the most specific for SLE?
becoz they are mainly seen in patients with lupus and their levels correlate with disease activity,
antibodies to dsDNA typically produce a ___ staining pattern on indirect immunofluorescence
peripheral or homogenous
methods in detecting ANA have been developed. These include ___
FANA or indirect immunofluorescent assay,
immunoperoxidase staining
enzyme linked immunoabsorbent assay
microsphere multiplex immunoassays
radioimmunoassay
immunodiffusion
immunoblottng (western blot)
dot blot
immunoelecterophoresis
microarray
what is the staining pattern in IIF of SLE
peripheral, homogenous,speckled, and nucleolar
if SLE only, the staining pattern is
peripheral
antibodies found in peripheral
anti DNA
not seen on HEp - 2
antibodies found in homogenous or diffuse staining pattern of IIF
anti - dna
anti - histone
anti DNP (nucleosome)
disease associated with homogenous pattern staining of IIF
RA
SLE
Miscellanous disorders (anti ss dna)
antibodies found in speckled pattern of IIF
anti sm & rnp
anti ro and la
anti - jo - 1 and mi 2
anti sci 70
diseases associated with speckled pattern staining in IIF
sle and ss
pm/dm
pss (systemic)
antibody found in centromere staining pattern in IIF
anti centromere
disease associated to centromere pattern in IIF
pss (crest)
anti body found in nucleolar staining in IIF
ANTI NUCLEOLAR
DISEASES found in nucleolar staining in IIF
sle and pss
the limited symptoms of scleroderma is referred to as
crest
transcribe crest
c
calcinosis
transcribe crest
r
raynaud’s phenomenon
transcribe crest
E
esophageal dysfunction
transcribe crest
S
sclerodactyly
transcribe crest
T
telangiectasis
refers to the calcium deposited in the skin
calcinosis
spasm of blood vessels in response to cold or stress
raynaud’s phenomenon
acid reflux and decrease in motility of esophagus
esophageal dysfunction
thickening and tightening of the skin in the fingers and hands
sclerodactyly
dilation of capillaries causing red marks on surface of the skin
telangiectasias
