Connective tissue diseases

Clinical information

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

Autoantibodies whose target antigens are located in the nucleus (so-called anti-nuclear antibodies, ANA) are important serological markers for CTD. Target antigens include, for example, nucleic acids, cell nuclear proteins and ribonuclear proteins. 

The frequency (prevalence) of anti-nuclear antibodies in inflammatory rheumatic diseases lies between 20 and 100 %. Therefore, differential ANA diagnostics is indispensable for the diagnosis 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. Positive and negative samples produce a large signal difference. In the microscopic evaluation it is possible to establish precisely how an indicator dye (generally fluorescein) is distributed in the tissue or the cells. A typical fluorescence pattern is produced for every bound autoantibody, depending on the location of the individual autoantigens.

The first international consensus on standardised nomenclature of HEp-2 cell patterns in indirect immunofluorescence (ICAP, www.anapatterns.org) defined fourteen nuclear patterns and nine cytoplasmic patterns which are relevant for the diagnosis of various autoimmune diseases.

Furthermore, the consensus recommends that autoantibodies detected in indirect immunofluorescence be confirmed by additional specific tests (e.g. ELISA, line blot). The exclusive use of these monospecific test methods is inadequate for the determination of autoantibodies against cell nuclei, as not all relevant antigens are available in a purified form as yet. Thus, the corresponding ANA can only be detected by IIFT.

Selected Products

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

human
IIFT
ANA Mosaic 2 EUROPattern
cell nuclei (ANA), EUROPattern
mitochondria (AMA)
2 BIOCHIPs per field:
HEp-2 cells
kidney

human
rat
IIFT
cell nuclei (ANA)
mitochondria (AMA)
HEp-2 cells
kidney
(2 BIOCHIPs per field)
human
rat
IIFT
cell nuclei (ANA) EUROPattern
cell nuclei (ANA)
HEp-20-10 cells
liver
(2 BIOCHIPs per field)
human
monkey
IIFT
cell nuclei
(ANA global test)
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
EUROPLUS ANA Mosaic 20A
cell nuclei (ANA)
cell nuclei (ANA)
SS-A + SS-B
ribosomal P-proteins + Jo-1
4 BIOCHIPs per field:
HEp-20-10 cells
liver
SS-A+SS-B BIOCHIPs
rib. P-prot.+Jo-1 BIOCHIPs

human
monkey
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)
cell nuclei (ANA)
HEp-20-10 cells
liver
(2 BIOCHIPs per field)
human
rat
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
Myositis Profile
(Mi-2, Ku, PM-Scl, Jo-1, PL-7,
PL-12, Ro-52 separately)
EUROLINE
EUROLINE
positive control serum: IgG, human, 100x concentrated
for DL 1530-X G
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
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
ELISA
histones
antigen-coated
microplate wells
native, highly purified histones,
bovine thymus
IIFT
antibodies against cell nuclei
(ANA control), homogeneous pattern
IIFT
test performance control
(ANA homogeneous, IgG)
IIFT
antibodies against cell nuclei (ANA control), homogeneous pattern,
control serum with indication of titer, reference serum W 1064 (66/233)
IIFT
antibodies against cell nuclei,
aliquots from positive control for ANA diagnostics
(20 different fluorescence patterns)
IIFT
antibodies against cell nuclei (ANA control),
homogeneous pattern, control serum with indication of titer
IIFT
antibodies against cell nuclei (ANA control),
homogeneous pattern, for EUROPattern
ELISA
double-stranded DNA
(dsDNA)
antigen-coated
microplate wells
highly purified genomic
double-stranded DNA
RIA
double-stranded DNA (dsDNA)
precipitation
labelled antigen
plasmid DNA
IIFT
antibodies against dsDNA
(dsDNA ab control)
IIFT
dsDNA
flagellates
Crithidia luciliae
IIFT
dsDNA
EUROPattern
flagellates
Crithidia luciliae
ELISA
dsDNA-NcX
antigen-coated
microplate wells
highly purified genomic
double-stranded DNA,
complexed with nucleosomes
IIFT
dsDNA (sensitive)
flagellates
Crithidia luciliae
IIFT
dsDNA (sensitive)
EUROPattern
flagellates
Crithidia luciliae
IIFT
antibodies against dsDNA (dsDNA ab control)
control serum with indication of titer
ELISA
nucleosomes
antigen-coated
microplate wells
native, nucleosomes,
calf thymus
ELISA
single-stranded DNA
(ssDNA)
antigen-coated
microplate wells
heat-denatured ssDNA
IIFT
antibodies against cell nuclei
(ANA control), nucleolar pattern
ELISA
PM-Scl
antigen-coated
microplate wells
recombinant,
expression in E. coli
IIFT
antibodies against cell nuclei
(ANA control), granular pattern
EUROLINE
positive control serum: IgG, human, 100x concentrated
for DL 1590-X G
ELISA
Anti-ENA ProfilePlus 1 (nRNP/Sm,
Sm, SS-A, SS-B, Scl-70, Jo-1 separately)
antigen-coated
microplate wells
see individual ELISA
EUROLINE
Anti-ENA ProfilePlus 1
(nRNP/Sm, Sm, SS-A, Ro-52, SS-B,
Scl-70, Jo-1 separately)
EUROLINE
EUROASSAY
Anti-ENA ProfilePlus
(nRNP/Sm, Sm, SS-A, SS-B, Scl-70,
Jo-1 separately)
EUROASSAY strip
with antigens
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
see individual ELISA
ELISA
Anti-ENA SLE Profile 2 (dsDNA, histones,
nucleosomes, nRNP/Sm, Sm,
SS-A, SS-B, Scl-70 separately)
antigen-coated
microplate wells
see individual ELISA
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
see individual ELISA
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
see individual ELISA
EUROASSAY
Anti-ENA ProfilePlus with AMA M2
(nRNP/Sm, Sm, SS-A, SS-B, Scl-70,
Jo-1, AMA M2 separately)
EUROASSAY strip
with antigens
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
EUROASSAY
Anti-ENA ProfilePlus with Ro-52
(nRNP/Sm, Sm, SS-A, SS-B, Scl-70,
Jo-1, Ro-52 separately)
EUROASSAY strip
with antigens
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
EUROASSAY
Anti-ENA ProfilePlus with ribosomal P-proteins
(nRNP/Sm, Sm, SS-A, SS-B, Scl-70,
Jo-1, ribosomal P-proteins separately)
EUROASSAY strip
with antigens
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
Mi2 alpha, Mi2 beta, Ku, DFS70
EUROLINE
EUROLINE
Cytoplasm profile
(AMA M2, M2-3E, ribosomal P-proteins, Jo-1
SRP, PL-7, PL-12, EJ, OJ, Ro-52 separately)
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
see individual ELISA
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
see individual ELISA
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
see individual ELISA
IIFT
antibodies against U1-nRNP
ELISA
nRNP/Sm
antigen-coated
microplate wells
native, U1-nRNP, incl. 70K, A, C, B, B',
D, E, F, G, calf thymus
IIFT
antibodies against Sm
ELISA
Sm
antigen-coated
microplate wells
native, U2-, U4-, U5-, U6-nRNP, incl. B, B',
D, E, F, G, calf thymus
IIFT
antibodies against SS-A (Ro)
ELISA
SS-A (Ro)
antigen-coated
microplate wells
native, SS-A-specific proteins (60 kDa),
calf thymus
IIFT
antibodies against SS-B (La)
ELISA
SS-B (La)
antigen-coated
microplate wells
native, SS-B-specific protein (48 kDa),
calf thymus
IIFT
antibodies against Scl-70
ELISA
Scl-70
antigen-coated
microplate wells
native, DNA topoisomerase I,
calf thymus
IIFT
antibodies against cell nuclei
(ANA control), pattern DFS70
ELISA
DFS70
antigen-coated
microplate wells
DFS70
recombinant
EUROLINE
EUROLINE Anti-DFS70
EUROLINE
IIFT
antibodies against PCNA (cyclin I)
IIFT
antibodies against mitosin (cyclin II)
IIFT
antibodies against Sp100
(nuclear granula, nuclear dots; associated with PBC)
IIFT
antibodies against Ku
IIFT
antibodies against nuclear membrane
IIFT
antibodies against cell nuclei
(ANA control), pattern centromeres
ELISA
centromeres
antigen-coated
microplate wells
recombinant centromere protein B,
expression with baculovirus vector
in insect cells
IIFT
antibodies against spindle fibers (MSA-2)
IIFT
antibodies against ribosomal P-proteins
ELISA
ribosomal P-proteins
antigen-coated
microplate wells
native, P0/P1/P2 of the 60S ribosomal
subunit, calf thymus
IIFT
antibodies against Golgi apparatus
IIFT
antibodies against histidyl-tRNA synthetase
(Jo-1 ab control)
ELISA
Jo-1
antigen-coated
microplate wells
native, histidyl-tRNA synthetase,
calf thymus
ELISA
cN-1A (Mup44, NT5C1A)
antigen-coated
microplate wells
recombinant, expression in E. coli
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