immuno concepts Colorzyme ANA Für Professionellen Gebrauch Seite 6

MITOTIC CELL (Metaphase)
trol), the mitotic cells will not show chromosome staining
and may be difficult to see.
USE OF MITOTIC CELLS
Discerning Speckled vs. Homogeneous Antibody: A
fine speckled pattern of staining is sometimes difficult
to differentiate from homogeneous staining. If the pat-
tern is homogeneous, there will be solid staining of the
chromosomes of the mitotic cells. If the pattern is strictly
speckled, the region outside of the chromosomes will
show a fine speckled reaction.
CELLULE MITOTIQUE
NOTE: If fine speckling of the entire mitotic cell occurs
along with solid staining of the chromosome region,
(Métaphase)
it is highly probable that two or more antibodies are
present. Report the screening dilution as speckled/
homogeneous and titer each antibody to endpoint.
Peripheral vs. Nuclear Membrane Antibody: Antibody
that shows a peripheral pattern is generally associated
with DNA/DNP nuclear antigens. High titers of these
antibodies are suggestive of SLE. In substrates that do
not include mitotic cells, the peripheral pattern can be
difficult to distinguish from nuclear membrane antibody.
By using Immuno Concepts' mitotic cells, these patterns
can be differentiated, because the chromosome
region of the mitotic cells will be stained intensely in a
peripheral pattern, but will not be stained by nuclear
CELLULA MITOTICA
membrane antibody. This distinction is clinically impor-
tant because nuclear membrane antibody does not
(Metafase)
have DNA/DNP specificity and is not associated with
SLE (35).
Anti-Centromere Antibody (ACA) vs. Atypical speckled
antibody resembling Centromere: In order to verify
anti-centromere antibody, the chromosome region
of the mitotic cells should stain brightly with discrete
speckles. If the chromosome region does not stain,
the antibody has neither kinetochore specificity nor as-
sociation with the CREST variant of scleroderma (36). If
the chromosome region does not stain, the antibody is
not anti-centromere, and should be reported as "atypi-
cal speckled."
CÉLULA MITÓTICA
CYTOPLASMIC STAINING
(Metafase)
Although autoantibodies to cytoplasmic antigens
are not commonly associated with connective tissue
disease, these antibodies may be detected using
epithelial cell culture substrates (37). Mitochondrial and
smooth muscle antibodies are the two most commonly
detected antibodies and are generally associated
with mononucleosis, chronic active hepatitis, and liver
disease (38- 39). Using the HEp-2 cell substrate, smooth
muscle antibody has also been demonstrated in pa-
tients with warts (40).
Anti-Mitochondrial Antibody (AMA): Discrete speckles
concentrated in the perinuclear region of the cell and
extended in lower density to the outer regions of the
MITOTISCHE ZELLE
cytoplasm. This should be distinguished from anti-Golgi
antibody, which generally stains only one side of the
(Metaphase)
perinuclear region, and from anti-ribosomal antibody,
which demonstrates finer speckles with a strandlike
appearance consistent with the location of the endo-
plasmic reticulum within the cell.
NOTE: Perinuclear speckles can most easily be dis-
tinguished from peripheral nuclear staining by noting
that the mitochondrial speckles form an interrupted
speckled staining around the outside of the nuclear
membrane, while peripheral sera form a solid smooth
staining inside the nuclear membrane.
REPORT SERA AS NEGATIVE FOR ANTINUCLEAR ANTIBOD-
IES AND VERIFY POSITIVE FOR ANTIMITOCHONDRIAL
MITOTISK CELL
ANTIBODY ON AMA SPECIFIC SUBSTRATE.
(Metafas)
Anti-Smooth Muscle Antibody (ASMA): Very fine
fibrous staining over the entire cytoplasm of cells with
Chromosome Region
Cytoplasm/
Nucleoplasm
Région chromosomique
Cytoplasme /
Nucléoplasme
La Zona Cromosomica
Citoplasma /
Nucleoplasma
Región Cromosómica
Citoplasma /
Nucleoplasma
Chromosomenbereich
Zytoplasma /
Nukleoplasma
ZELLE AUSSERHALB DER MITOSEPHASE
Kromosomområde
Cytoplasma /
Nukleoplasma
NON-MITOTIC CELL (Interphase)
a "spiderweb" appearance. Unlike mitochondrial
antibody, smooth muscle antibody staining is uniform
over the entire cytoplasm and may also extend over
the nucleus. Mitotic cells generally show large discrete
speckles outside the chromosome region. Smooth
muscle antibody has been shown to have a high speci-
ficity to actin (41-42).
REPORT SERA AS NEGATIVE FOR ANTINUCLEAR ANTI-
BODY AND VERIFY POSITIVE FOR ANTI-SMOOTH MUSCLE
ANTIBODY ON ASMA SPECIFIC SUBSTRATE.
CELLULE NON MITOTIQUE
LIMITATIONS OF THE TEST
(Interphase)
1. Diagnosis cannot be made on the basis of anti-
nuclear antibody detection alone. The physician
must interpret these results in conjunction with the
patient's history and symptoms, the physical find-
ings, and other diagnostic procedures.
2. Treatment should not be initiated on the sole basis
of a positive test for antinuclear antibodies. Clini-
cal indications, other laboratory findings, and the
physician's clinical impression must be considered
before any treatment is initiated.
3. Certain drugs, including procainamide and
hydralazine, may induce a lupus erythemato-
CELLULA NON IN MITOSI
sus-like disease (43). Patients with drug-induced
LE may demonstrate positive homogeneous
(Interfase)
or homogeneous/peripheral ANAs commonly
directed against nuclear histones (44).
4. A small percentage of patients with SLE may not
demonstrate ANAs by indirect immunoenzyme but
may have ANAs by other techniques (45).
5. Although a high-titered ANA may be highly sugges-
tive of connective tissue disease, it should not be
considered diagnostic but rather viewed as a part
of the overall clinical history of a patient.
6. Staining patterns often change with progressive
titration of sera. This phenomenon is generally
CÉLULA NO MITÓTICA
due to the presence of more than one nuclear
antibody.
(Interfase)
7. Positive ANAs are also seen in a small percent-
age of patients with infectious and/or neoplastic
diseases (9).
EXPECTED VALUES
In a large university medical center, using HEp-2 cell
ANA substrate, the following data were generated over
a two-year period (46). Table 1.
PERFORMANCE CHARACTERISTICS
A comparative study has shown the Immuno Concepts
Colorzyme® ANA Test System to be equivalent to the
Immuno Concepts indirect immunofluorescent ANA
(Interphase)
test. Fifty serum samples were screened and titered on
both systems. These samples included autoantibodies
commonly detected with the HEp-2 substrate, uncom-
mon and atypical patterns, and borderline negative
sera. The enzyme results correlated 98% with the
Immuno Concepts immunofluorescent ANA for positive,
negative, and mixed pattern sera tested. (46).
The Immuno Concepts Fluorescent ANA test systemm
was previously evaluated in comparison with two other
fluorescent antibody tests in commercial distribution
(46). The study employed 97 serum samples from
normal individuals as well as from diagnoses including
systemic lupus erythematosus (SLE), mixed connective
ICKE-MITOTISK CELL
tissue disease (MCTD), Raynaud's-progressive systemic
sclerosis-CREST variant (PSS-CREST), rheumatoid arthritis
(Interfas)
(RA), juvenile rheumatoid arthritis (JRA), as well as other
connective tissue disease. Sera were tested at the rec-
6
Cytoplasm
Nuclear Membrane
Nucleoplasm
Nucleoli
Cytoplasme
Membrane Nucléaire
Nucléoplasme
Nucléoles
Citoplasma
Membrana Nucleare
Nucleoplasma
Nucleoli
Citoplasma
Membrana Nuclear
Núcleo
Nucleólos
Zytoplasma
Zellkermembran
Nukleoplasma
Nucleoi
Cytoplasma
Nukleärt Membran
Nukleoplasma
Nukleol