01/04/2010 Pathology Associates Of Lexington, P.A.

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All ANA tests must be performed on diluted serum so as to dilute out lots of nonspecific "reactivity" (standard is a 1:10
dilution of serum). The classical "ANA" is a fluorescent auto-immune antibody (Ab) screening test visually performed
using a fluorescent microscope (so, sometimes called F-ANA) and is an "intellect intensive" test. F-ANA usually looks
for ANAs in the IgG class (not in IgA or IgM). The substrate which provides the nuclei is either made of histological
sections of tissue (cross-sections of nuclei) or cell culture monolayer sheets which have whole nuclei (such as HEp-
2). Depending on source or method of substrate production, cells may contain typical or scant amounts of the various
antigens. [ANA] [autoimmune ] [muscle Bx & comprehensive muscle lab & path link]

Many labs use the automated, more "chemical", non-visual (ELISA) test methods for ANA, and such methods do not
reveal visual patterns.
specific antibody
antigen HEp-2 ANA pattern disease associations
designation 5
nucleic acid 1. deoxyribonucleoprotein 1. anti-DNP 1. homo/peripheral 1. hi titer, SLE (95%
antigens 2. native double stranded 2. anti-DNA 2. homo/peripheral specific for SLE4);
deoxyribonucleic acid 3. anti-RNA 3. ? low titer in RA or
(confirm with Crithidia 4. anti- 4. nucleolar drug induced SLE
luciliae positivity) nucleolar 2. hi titer SLE, CAH;
3. ribonucleic acid low titer in JCP
4. 4-6s RNA 3. SLE
4. highly associated
with PSS &
Raynaud's

histone 1. histone 1. anti-histone 1. homo/peripheral 30% of SLE & 90% of

nuclear 2. histone (H3?) 2. anti-histone 2. variable large procainamide-induced
protein (H3?) speckled LE

non-histone 1. Smith 1. anti-Sm (an 1. coarse nuclear 1. highly specific

nuclear 2. ribonucleoprotein [U1- anti-ENA) speckled4 for SLE but
protein snRNP] 2. anti-nRNP 2. coarse nuclear present in only
3. Sjogrens syn. A [same (an anti- 20%; especially in
speckled4
as Ro] ENA) AAs4
3. coarse to fine
4. Sjogrens syn. B [same 3. anti-SS-A 2. M CTD, SLE, &
speckled & with
as La or Ha] [same as scleroderma
some weak
5. MA-1 anti-Ro] (an 3. 95% of Sjogren's
cytoplasmic
6. scleroderma 70 anti-ENA)
granules4 sicca complex4;
7. prolif. cell nuc. ag. 4. anti-SS-B
4. uniform fine neonatal maternal
[same as
speckled SLE assoc. heart
anti-La &
block; & 66% of
anti-Ha] (an nuclear4
"ANA-negative"
anti-ENA) 5. beaded
SLE.
5. anti-MA-1 peripheral5 4. Sjogren's sicca
6. anti-Scl-70 6. speckled complex & 20% of
(an anti- 7. variegated SLE
ENA) speckled (30- 5. severe SLE
7. anti-PCNA 60% of cells) 6. 15-30% of PSS
7. SLE with diffuse
proliferative
glomerulonephritis

other 1. chromosomal 1. anti- 1. discrete 1. CREST & CREST-

specificities centromere centromere speckled PBC
2. mitotic spindle (ACA) 2. mitotic spindle 2. non-specific
apparatus (NuMA; 2. anti-MSA 3. irregular connective tissue
MSA-1) 3. anti-nuclear speckled disorder
3. nuclear matrix matrix 4. ? 3. ?
4. Mi-1 4. anti-Mi-1 5. fine speckled 4. dermatomyositis ?
5. Jo-1 5. anti-Jo-1 (& cytoplasmic 5. 30% of
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01/04/2010 Pathology Associates Of Lexington, P.A.
6. Ku some other 6. ? polymyositis
7. Su (systems A,B,C) anti-tRNA 7. ? 6. scleroderma &
8. PM-1 symthetases) 8. ? polymyositis
9. SL (an anti- 9. ?speckled overlap
10. RANA ENA) 10. ? 7. ?
6. anti-Ku 8. 40-50% of
7. anti-systems polymyositis
(A,B,C) 9. SS-SLE overlap
8. anti-PM-1 10. hi titer likely RA
9. anti-SL
10. anti-RANA

cytoplasmic 1. Actin 1. anti-actin 1. Fibrillar 1. CAH

antigens 2. Centrioles 2. anti- 2. Large bodies 2. ?
3. Golgi centriolar 3. Perinuclear 3. SS; SLE
4. Lysosomes 3. anti-Golgi strands 4. SLE
5. Mitochondria 4. anti- 4. Round granules 5. PBC
6. Ribosomes/ribonuc. lysosomal 5. Irregular 6. SLE
protein 5. anti- granules 7. in many diseases
7. Vimentin mitochondrial 6. Fine cyto.
6. anti- speckles
ribosomal 7. Large fibrils
7. Anti-vimentin

term4 pattern description disease associations

uniform diffuse pos. of interphase common in SLE, drug induced

homogeneous nuclear
hom. nuclear with rim
nuclear membranous-linear
nuclear membrane pores
large speckled or nuclear matrix
coarse speckled
fine speckled
pleomorphic
nuclear material; with metaphase lupus, RA, juvenile chronic
concentration of positivity ain arthritis, and in systemic sclerosis
chromosome region & neg. (with intense fine speckled
cytoplasm patterns)
as above, with greater intensity
common in SLE, especially when
at the outer region of the
active
interphase nucleus
fine, linear positivity of the inner
in unusual mixed chronic
and outer nuclear membranes
autoimmune disorders, usually
with less intense homogeneous
including chronic hepatitis and
staining of the entire nucleus and
variably associated with vasculitis,
some nucleolar staining;
thrombocytopenia and Raynaud's
metaphase chromatin negative
phenomenon
and cytoplasmic positive
nuclear membrane shows a
granular/speckled staining of
interphase nuclei; in metaphase,
PBC; polymyositis
membrane fragments into small
particles causing homogeneous
chromosomal staining
variable large speckles in a
sponge-like nuclear network in MCTD, SLE, & other chronic
interphase; metaphase, rheumatic diseases
chromatin negative
dense intermediate sized
particles together with large
SLE & SLE overlap cases
speckles; metaphase chromatin
negative
fine to discrete speckled
interphase uniformly distributed; SLE, SS, SCLE, & scleroderma
metaphase, chromatin negative
varying fine to coarse speckling
1-3% of SLE with
in 30-60% of interphase cells;
glomerulonephritis and
positive or negative metaphase
lymphoproliferative disorders
chromatin

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centromere or discrete speckled
few nuclear dots
multiple nuclear dots
nucleolar homogeneous
nucleolar clumpy pattern
nucleolar speckled pattern
nucleolar speckled with mitotic
dots
tubulin
centrosome (centriole)
nuclear mitotic apparatus
midbody (MSA-2)
mitotic spindle antigen (MSA-3)
fine speckled pattern (or Jo-1
pattern)
ribosomal pattern
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Pathology Associates Of Lexington, P.A.
60% of CREST; patients with
primary Raynaud's who are
40-60 discrete fine speckles;
centromere antibody positive
interphase chromatin positive
frequently develop limited
scleroderma
0-6 discrete round dots in
interphase, often adjacent to the CAH, viral, PBC
nucleolus
in interphase: dots are peripheral
in metaphase and not in
6% of PBC and often with SS and
chromatin (allowing it to be
less frequent in SLE
distinguished from the
centromere pattern)
50% of cases of polymyositis-
scleroderma overlap; high
entire nucleolar staining
frequency in primary pulmonary
hypertension with scleroderma
hi specificity for PSS, 5% of cases,
brightly clustered large granules especially with skeletal muscle
in the nucleoli disease and pulmonary
hypertension
nucleoli positive by way of 10-20 hi specificity for PSS, 30% of
irregular dots cases
many speckles in nucleoli of
interphase cells; speckles in scleroderma with Raynaud's
metaphase chromatin distinguish phenomenon
it from "nucleolar speckled"
staining of the mitotic spindle with
higher titers in mono, alcoholic
pole to pole and kinetochore to
cirrhosis, Hashimoto's,
pole fibers visible; and, fine
thyrotoxicosis, chronic parasitic
filamentous staining in cytoplasm,
infections, and other chronic
which condenses where two cells
diseases
abutt.
metaphase mitotic spindles
display discrete spots at each non-specific rheumatic diseases;
pole and one or two bright spots some chronic post viral syndromes
in cytoplasm of interphase
positive spindle poles in
metaphase & fine speckled non-specific and seen in SLE, SS,
nuclear staining with negative CREST, MCTD, & polyarthritis
nucleoli in interphase
equatorial plane area positivity of
metaphase nuclei or at the systemic sclerosis & Raynaud's
constriction points linking to phenomenon
daughter cells during telophase
a fine, dense nuclear speckled
pattern in some interphase cells
with speckles aligned along the seen in some patients with
spindle in prophase and respiratory tract tumors
metaphase; telophase cells are
negative
fine cytoplasmic granules
20-40% of patients with
condensed around the nucleus,
aggressive polymyositis; interstitial
which diminish towards the
lung disease; arthralgia
periphery
prominent fine, dense granular to
homogeneous cytoplasmic
associated with SLE patients with
staining with perinuclear
neuropsychiatric symptoms and
accentuation plus nucleolar
renal disease, often with absence
staining but no nuclear staining
of dsDNA antibodies.
(similar to Jo-1 but speckles
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finer)
numerous coarse
common in PBC; about 40% of
granular/filamentous cytoplasmic
mitochondrial scleroderma; sometimes in
speckles around the nucleus and
overlap syndromes
throughout cytoplasm
fine speckled to granular
cytoplasmic pattern with no 4% of patients with
signal recognition particle
nuclear or nucleolar staining polymyositis/dermatomyositis
(similar to ribosomal pattern)
large granules condensed in the drug-induced hepatitis and
cytoplasm and more pronounced anticonvulsant hypersensitivity;
endoplasmic reticulum
at one side of the nucleus (seen sometimes after infections as
as LKM-1 on rodent tissues protein production is stimulated
large irregular speckles uncommon pattern seen in SLE
lysosomal pattern distributed throughout the but without useful clinical
cytoplasm associations
a moderately fine, uniform sized rare pattern found in patients with
granular staining of the poorly defined musculoskeletal
peroxisomes
cytoplasm in a random pattern; pains, thyroid disorders, and
no nuclear or chromatin staining polymyalgia rheumatica
speckled polar staining adjacent
uncommonly found with SLE, SS,
to one part of the nucleus &
golgi complex and other chronic systemic
composed of irregular large
rheumatic disease
granules
fascicles of fine fluorescent fibers
chronic active hepatitis, where it
crossing the cytoplasm and
actin (smooth muscle) may be associated with a nuclear
frequently as part of a mixed
speckled pattern and nuclear dots
pattern with other antibodies
reticular strand pattern
30-80% of patients with
throughout cytoplasm and
rheumatoid arthritis; MCTD; CAH;
concentrated around the nucleus
cytokeratin SCC of lung; Crohn's disease; and
and along cytoplasmic extensions
many other diseases, as well as
(similar to vimentin but with fewer
normals
cytoplasmic whorls)
found in a wide variety of patients
stress fibers running the length
with rheumatoid arthritis, MCTD,
of the cells and (like actin)
tropomyosin CAH, SCC of lung, Crohn's
appear to hold the cells under
disease, and many other diseases
tension
and normals
abundant fine radiating fibers found in a variety of diseases
through the cytoplasm and such as chronic rheumatic and
making connections with nuclear liver diseases, SLE, cancer and
vimentin
and plasma membrane in hematological disorders and also
complex, reticular, whorled in chronic inflammatory disorders
patterns such as Crohn's disease
short cable-like extensions non-specific association of a
vinculin adjacent to the nuclear and variety of autoimmune and chronic
cytoplasmic membranes diseases
linear and granular cytoplasmic
desmin pattern with some nuclear ?
component
short cable-like extensions of cell
Cytoskeletal anchoring proteins cytoplasm to other cells or to sometimes in systemic sclerosis
extracellular matrix

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