Connective tissue diseases

Clinical information

Connective tissue diseases (CTD) are rheumatic diseases. They include, for example, systemic lupus erythematosus, systemic sclerosis, Sjögren's syndrome, myositis syndromes or Sharp syndrome.

Autoantibodies against cellular components, commonly called anti-nuclear antibodies (ANA), are important serological markers for CTD. Target antigens include nucleic acids, cell nuclear proteins and ribonuclear proteins as well as components of the cytoplasm. The frequency (prevalence) of ANA in inflammatory rheumatic diseases lies between 20% and 100%. Differential ANA diagnostics is therefore indispensable for the identification of individual rheumatic diseases and their differentiation from other autoimmune diseases.

Diagnostics

The gold standard for the determination of ANA is the indirect immunofluorescence test (IIFT) with human epithelial cells (HEp-2), which is known for its high sensitivity and specificity. In the microscopic evaluation it is possible to establish precisely how an indicator dye (generally fluorescein) is distributed in the cells. Depending on the location of the individual autoantigens, a typical fluorescence pattern for every type of autoantibody bound will appear.

The revised international consensus on a standardised nomenclature of HEp-2 cell patterns in indirect immunofluorescence (ICAP, www.anapatterns.org) defines 15 nuclear, 9 cytoplasmic and 5 mitotic patterns which are relevant for the diagnostics of various autoimmune diseases.

Furthermore, ICAP recommends that IIFT results be confirmed by additional specific antibody tests (e.g. ELISA, line blot, ChLIA). However, the exclusive use of these monospecific test methods is insufficient for the determination of autoantibodies against cell nuclei, as not all relevant antigens are available yet in purified form. Thus, the corresponding ANAs can only be detected by IIFT.

Files

Strategy for the Determination of ANA and Cytoplasm Components

ELISA test systems for the determination of antibodies against nuclear and cytoplasmic antigens

EUROLINE Nuclear and Cytoplasmic Antigen Profiles

EUROLINE myositis profiles

EUROLINE systemic sclerosis profiles

Chemiluminescent immunoassays for rheumatic disease diagnostics

 

Myositis diagnostics
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Techniques

Method
Parameter
Substrate
Species
IIFT
cell nuclei
(ANA global test)
HEp-2 cells
liver
(2 BIOCHIPs per field)
human
monkey
IIFT
cell nuclei (ANA) EUROPattern
cell nuclei (ANA)
HEp-2 cells
liver
(2 BIOCHIPs per field)
human
monkey
IIFT
cell nuclei (ANA)
mitochondria (AMA)
HEp-2 cells
kidney
(2 BIOCHIPs per field)
human
rat
IIFT
cell nuclei
(ANA global test)
HEp-20-10 cells
liver
(2 BIOCHIPs per field)
human
monkey
IIFT
cell nuclei (ANA) EUROPattern
cell nuclei (ANA)
HEp-20-10 cells
liver
(2 BIOCHIPs per field)
human
monkey
IIFT
EUROPLUS ANA Mosaic 10A
cell nuclei (ANA)
SS-A + SS-B
2 BIOCHIPs per field:
HEp-20-10 cells
SS-A+SS-B BIOCHIPs

human
IIFT
cell nuclei (ANA)
mitochondria (AMA)
HEp-20-10 cells
kidney
(2 BIOCHIPs per field)
human
rat
IIFT
cell nuclei (ANA) EUROPattern
mitochondria (AMA)
HEp-20-10 cells
kidney
(2 BIOCHIPs per field)
human
rat
IIFT
EUROPLUS ANA Mosaic 22A
cell nuclei (ANA)
cell nuclei (ANA)
nRNP/Sm + Sm + SS-A
SS-B + Scl-70 + Jo-1
4 BIOCHIPs per field:
HEp-20-10 cells
liver
nRNP/Sm+Sm+SS-A BIOCHIPs
SS-B+Scl-70+Jo-1 BIOCHIPs

human
monkey
IIFT
cell nuclei (ANA)
HEp-2 cells
human
IIFT
cell nuclei (ANA)
EUROPattern
HEp-2 cells
human
IIFT
cell nuclei (ANA)
HEp-20-10 cells
human
IIFT
cell nuclei (ANA)
EUROPattern
HEp-20-10 cells
human
EUROLINE
positive control serum: IgG, human, 100x concentrated
for DL 1530-X G
EUROLINE
Myositis Profile
(Mi-2, Ku, PM-Scl, Jo-1, PL-7,
PL-12, Ro-52 separately)
EUROLINE
EUROLINE
Myositis Profile 3
(Mi-2, Ku, PM-Scl100, PM-Scl75, Jo-1, SRP,
PL-7, PL-12, EJ, OJ, Ro-52 separately)
EUROLINE
EUROLINE
Autoimmune Inflammatory Myopathies 16 Ag
(Mi-2 alpha, Mi-2 beta, TIF1g, MDA5, NXP2, SAE1, Ku, PM-Scl100,
PM-Scl75, Jo-1, SRP, PL-7, PL-12, EJ, OJ, Ro-52 separately)
EUROLINE
EUROLINE
Autoimmune Inflammatory Myopathies 16 Ag et cN-1A
(Mi-2 alpha, Mi-2 beta, TIF1g, MDA5, NXP2, SAE1, Ku,
PM-Scl100, PM-Scl75, Jo-1, SRP, PL-7, PL-12, EJ, OJ,
Ro-52, cN-1A separately)
EUROLINE
EUROLINE
Autoimmune Inflammatory Myopathies 20 Ag
( Mi-2 alpha, Mi-2 beta, TIF1g, MDA5, NXP2, SAE1,
Ku, PM-Scl100, PM-Scl75, Jo-1, SRP, PL-7, PL-12, EJ, OJ,
Ro-52, cN-1A, Ha, Ks, Zo)
EUROLINE
EUROLINE
positive control serum: IgG, human, 100x concentrated
for DL 1532 G
EUROLINE
Systemic Sclerosis Profile (Nucleoli)
(Scl-70, CENP A, CENP B, RP11, RP155,
fibrillarin, NOR90, Th/To, PM-Scl100, PM-Scl75,
Ku, PDGFR, Ro-52 separately)
EUROLINE
EUROLINE
Systemic Sclerosis Profile 2
(Scl-70, CenpA, Cenp B, RP11, RP155,
Fibrillarin, NOR90, Th/To, PM-Scl100, PM-Scl75,
Ku, NVL und Ro-52)
EUROLINE
ELISA
histones
antigen-coated
microplate wells
IIFT
antibodies against cell nuclei
(ANA control), homogeneous pattern
ChLIA
IDS dsDNA IgG

antigenic coated magnetic particles
ELISA
double-stranded DNA
(dsDNA)
antigen-coated
microplate wells
IIFT
dsDNA
EUROPattern
flagellates
Crithidia luciliae
IIFT
dsDNA
flagellates
Crithidia luciliae
IIFT
antibodies against dsDNA
(dsDNA ab control)
ELISA
dsDNA-NcX
antigen-coated
microplate wells
IIFT
dsDNA (sensitive)
EUROPattern
flagellates
Crithidia luciliae
IIFT
dsDNA (sensitive)
flagellates
Crithidia luciliae
ELISA
nucleosomes
antigen-coated
microplate wells
ELISA
single-stranded DNA
(ssDNA)
antigen-coated
microplate wells
ELISA
PM-Scl
antigen-coated
microplate wells
ChLIA
IDS ENA Screen

antigenic coated magnetic particles
EUROLINE
positive control serum: IgG, human, 100x concentrated
for DL 1590-X G
ChLIA
IDS ANA Screen

antigenic coated magnetic particles
ChLIA
IDS ANA Control Set
3 x 1.5mL Control 1/2
EUROASSAY
Anti-ENA ProfilePlus
(nRNP/Sm, Sm, SS-A, SS-B, Scl-70,
Jo-1 separately)
EUROASSAY strip
with antigens
ELISA
Anti-ENA ProfilePlus 1 (nRNP/Sm,
Sm, SS-A, SS-B, Scl-70, Jo-1 separately)
antigen-coated
microplate wells
EUROLINE
Anti-ENA ProfilePlus 1
(nRNP/Sm, Sm, SS-A, Ro-52, SS-B,
Scl-70, Jo-1 separately)
EUROLINE
ELISA
ANA screen 11 (antigen mixture: PCNA,
PM-Scl, ribosomal P-proteins, nRNP/Sm,
Sm, SS-A, Ro-52, SS-B, Scl-70, Jo-1,
centromeres)
antigen-coated
microplate wells
ELISA
Anti-ENA SLE Profile 2 (dsDNA, histones,
nucleosomes, nRNP/Sm, Sm,
SS-A, SS-B, Scl-70 separately)
antigen-coated
microplate wells
ELISA
ANA screen 9 (antigen mixture: PCNA,
PM-Scl, ribosomal P-proteins, nRNP/Sm,
Sm, SS-A, SS-B, Scl-70, Jo-1)
antigen-coated
microplate wells
ChLIA
IDS ANA Screen Control Set
3 x 1.5mL Control 1/2
ELISA
Anti-ENA ProfilePlus 2 (ribosomal
P-proteins, nRNP/Sm, Sm, SS-A, SS-B,
Scl-70, Jo-1, centromeres, separately)
antigen-coated
microplate wells
EUROLINE
Anti-ENA ProfilePlus 2
(nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70,
Jo-1, ribosomal P-proteins separately)
EUROLINE
EUROASSAY
Anti-ENA ProfilePlus with histones
(nRNP/Sm, Sm, SS-A, SS-B, Scl-70,
Jo-1, histones separately)
EUROASSAY strip
with antigens
EUROLINE
ANA Profile 23
(nucleosomes, dsDNA, histones, SS-A, Ro-52,
SS-B, nRNP/Sm, Sm, Mi-2 alpha, Mi-2 beta, Ku, CENP A,
CENP B, Sp100, PML, Scl-70, PM-Scl100, PM-Scl75,
RP11, RP155, gp210, PCNA, DFS70 separately)
EUROLINE
EUROLINE
ANA Profile 3
(nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70, PM-Scl,
Jo-1, CENP B, PCNA, dsDNA, nucleosomes,
histones, ribosomal P-proteins, AMA M2 separately)
EUROLINE
EUROLINE
ANA Profile 3 plus DFS70
(nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70, PM-Scl,
Jo-1, CENP B, PCNA, dsDNA, nucleosomes, histones,
ribosomal P-proteins, AMA M2, DFS70 separately)
EUROLINE
EUROLINE
ANA Profile et Mi-2 et Ku
(Mi-2, Ku, nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70,
PM-Scl, Jo-1, CENP B, PCNA, dsDNA, nucleosomes,
histones, ribosomal P-proteins, AMA M2 separately)
EUROLINE
EUROLINE
dsDNA, nucleosomes, histones, DFS70
EUROLINE
EUROLINE
ANA Profile et Mi-2, Ku, DFS70
(Mi-2, Ku, nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70,
PM-Scl100, Jo-1, CENP B, PCNA, dsDNA,
nucleosomes, histones, ribosomal P-proteins,
AMA M2, DFS70 separately)
EUROLINE
EUROLINE
positive control serum: IgG, human, 100x concentrated
for DL 1590-35 G
EUROLINE
Cytoplasm profile
(AMA M2, M2-3E, ribosomal P-proteins, Jo-1
SRP, PL-7, PL-12, EJ, OJ, Ro-52 separately)
EUROLINE
EUROLINE
Cytoplasm Profile 13 Ag
CAMA-M2, M2-3E, ribosomale P-Proteine,
Jo-1, SRP, PL-7, PL-12, EJ, OJ, Ro-52, Ha, Ks, Zo)
EUROLINE
EUROLINE
ANA Profile 5
(nRNP/Sm, Sm, RNP70, RNPA, RNPC, SS-A,
Ro-52, SS-B, Scl-70, PM-Scl, Jo-1, CENP B,
PCNA, dsDNA, nucleosomes, histones,
ribosomal P-proteins, AMA M2 separately)
EUROLINE
ELISA
Anti-ENA PoolPlus (antigen mixture:
nRNP/Sm, Sm, SS-A, SS-B, Scl-70, Jo-1)
antigen-coated
microplate wells
ELISA
ANA screen (antigen mixture: dsDNA,
histones, ribosomal P-proteins, nRNP/Sm,
Sm, SS-A, SS-B, Scl-70, Jo-1, centromeres)
antigen-coated
microplate wells
EUROLINE
ANA Profile 1
(nRNP/Sm, Sm, SS-A, Ro-52, SS-B, Scl-70,
Jo-1, CENP B, dsDNA, nucleosomes, histones,
ribosomal P-proteins separately)
EUROLINE
ELISA
Anti-ENA Pool (antigen mixture:
nRNP/Sm, Sm, SS-A, SS-B, Scl-70,
ribosomal P-proteins, Ro-52)
antigen-coated
microplate wells
ChLIA
IDS U1-snRNP

antigenic coated magnetic particles
ELISA
nRNP/Sm
antigen-coated
microplate wells
ChLIA
IDS Sm

antigenic coated magnetic particles
ELISA
Sm
antigen-coated
microplate wells
ChLIA
IDS SS-A/Ro

antigenic coated magnetic particles
ELISA
SS-A (Ro)
antigen-coated
microplate wells
ChLIA
IDS SS-A/Ro 52 kDa

antigenic coated magnetic particles
ChLIA
IDS SS-A/Ro 60 kDa

antigenic coated magnetic particles
ChLIA
IDS SS-B/La

antigenic coated magnetic particles
ELISA
SS-B (La)
antigen-coated
microplate wells
ChLIA
IDS Scl-70

antigenic coated magnetic particles
ELISA
Scl-70
antigen-coated
microplate wells
EUROLINE
positive control serum: IgG, human, 100x concentrated
for DL 159z-X G
ELISA
DFS70
antigen-coated
microplate wells
EUROLINE
EUROLINE Anti-DFS70
EUROLINE
ChLIA
IDS Centromere B

antigenic coated magnetic particles
ELISA
centromeres
antigen-coated
microplate wells
ELISA
ribosomal P-proteins
antigen-coated
microplate wells
ChLIA
IDS Jo-1

antigenic coated magnetic particles
ELISA
Jo-1
antigen-coated
microplate wells
ELISA
cN-1A (Mup44, NT5C1A)
antigen-coated
microplate wells
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