Antinuclear Antibodies (Anas) Antibodies To Extractable Nuclear Antigens (Anti-Enas)
Antinuclear Antibodies (Anas) Antibodies To Extractable Nuclear Antigens (Anti-Enas)
Antinuclear Antibodies (Anas) Antibodies To Extractable Nuclear Antigens (Anti-Enas)
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of
ELISA and IMMUNOBLOT kits are optimized and validated for detection of IgG
antinuclear antibodies in human serum and plasma
Determination of antinuclear antibodies is important for diagnosis of systemic autoimmune diseases. These organ non-
specific auto-antibodies are directed to intracellular antigens located mainly in the nucleus of the cell. Their detection
can indicate the presence of some systemic autoimmunopathologic process - especially: systemic lupus erythematodes
(SLE), Sjögren’s syndrome (SS), scleroderma, mixed connective tissue disease (MCTD), systemic sclerosis, polymyositis
and dermatomyositis.
An important group of antinuclear antibodies represent antibodies against ENA (extractable nuclear antigens: SS-A/Ro,
SS-B/La, Sm, RNP, Scl-70 and Jo-1). They are mainly ribonucleoproteins and nuclear enzymes.
Identification of single auto-antibody specificity is an important tool for differential diagnosis of systemic auto-immune
diseases. Antibodies against SS-A/Ro and SS-B/La often occur in patients with SS and SLE. They can be also found
in mothers of children with neonatal lupus and congenital heart block. Anti-Sm antibodies represent a highly specific
marker and one of diagnostic and classification criteria for SLE. Also anti-RNP antibodies (a part of Sm/RNP complex)
are often detected in patients with SLE. Presence of these antibodies is highly specific for MCTD (particularly when
anti-Sm antibodies are missing). Detection of anti-Jo-1 antibodies is significant for another group of organ non-specific
autoimmune diseases – myositides. Antibodies against antigen Scl-70 and centromere B are typical for diagnosis of
systemic sclerosis (particularly its progressive forms).
The group of antinuclear antibodies also includes antibodies against nucleic acids (ssDNA, dsDNA), complexes of nuclear
proteins (DNP, RNP) and histones.
The assay is based on a sandwich type of ELISA method. EIA ENA screen plus - Mixture of native and recombinant
antigens: SS-A/Ro52, SS-A/Ro60, SS-B/La, RNP A, RNP C,
RNP 68, SmB, SmD, Scl-70, Jo-1, Centromere B
EIA SS-A - Highly purified native antigen SS-A/Ro60
Sandwich ELISA
(60 kDa) and recombinant antigen SS-A/Ro52 (52 kDa)
EIA SS-A/Ro60 - Highly purified native antigen SS-A/Ro60
HRP
(60 kDa)
EIA SS-A/Ro52 - Recombinant antigen SS-A/Ro52 (52 kDa)
Anti-Hu IgG
EIA SS-B - Mixture of native and recombinant antigen
SS-B/La
EIA Sm - Mixture of recombinant antigen SmB and native
IgG
antigen SmD
EIA U1RNP - Mixture of recombinant antigens RNP A, RNP
C a RNP 68
Ag
Microtiter plate
EIA Scl-70 - Mixture of native and recombinant antigen
Scl-70
EIA Centromere - Mixture of recombinant antigens CENP B
a CENP A
SUMMARY PROTOCOL EIA Jo-1 - Recombinant antigen Jo-1
CLINICAL APPLICATION
Step Test steps
Blood donors (n = 227) - control group Patients with autoimmune disease (n = 143)
Sclerodermia 15 14 93%
Dermatomyositis 11 11 100%
Unclear systemic
33 29 88%
autoimmune disease
Immunoblot Immunoblot
Positive samples Blood donors
% reaction % reaction
(n = 380) (n = 227)
pos neg pos neg
Correlation Correlation
% reaction 80% 20% % reaction 4% 96%
90% 96%
60%
54%
50%
24% 21%
30%
14%
20%
3% 6% 9% 1% 1%
10%
0%
A
68
P0
A
La
0
B
2
0
Sm
N
o6
P
o5
P
l-7
P
D
N
N
EN
N
R
ds
R
R
Sc
R
C
P Reaction of substrate
and enzyme resulting BLOT-LINE ANA
in coloured insoluble product
E Confirmatory test for EIA ENA screen plus
Diferential diagnosis of systemic autoimmune diseases
Enzyme-conjugated secondary antibody by determination of specific ANAs
binding to primary antibody
IgG IgG
Start Start
Jo-1 Polymyositis Jo-1
PL-7
PL-12
Dermatimiositis CENP-B
PmScl Scl-70
CENP-A
CENP-B Systemic sclerosis
Scl-70 Ro52
polymyositis
Jo-1 Histidyl-tRNA synthetase dermatomyositis
increased risk of pulmonary fibrosis
ELISA IMMUNOBLOT
No. of No. of
Cat. No. Product Cat. No. Product
Tests Tests
ENA096 EIA ENA screen plus 96 ENAL20 BLOT-LINE ENA plus 20
Sm0096 EIA Sm 96
CONTACT
RQA RQA
R L R L
TE TE
IS
IS
LLOY ‘S REG
LLOY ‘S REG
D
V11/2017