immuno concepts Colorzyme ANA Für Professionellen Gebrauch Seite 4

†This slide is protected by one or more of the following
U.S. and foreign patents: 4387972; D-274261; D-273261;
0727046; Canadian patent 1171302 and other patents
pending.
†† This material is protected by one or more of the
following U.S. and foreign patents: 4615972; Canadian
patent 1231633. Italian patent 1177110 and other
patents pending.
NON-REACTIVE REAGENTS
PBS Buffer Powder: Catalog No. 1011. Phosphate-buff-
ered saline powder (0.01 M, pH 7.4 ± 0.2). Each pouch
contains sufficient buffer powder to make 1 liter. (One
pouch of buffer powder is supplied for every five slides
in complete test kits.)
Preparation: Dissolve one pouch of buffer powder in
1 liter of deionized or distilled water, cover, and store
refrigerated at 2-10°C for up to four weeks or until signs
of contamination or other visible changes occur.
Semi-Permanent Mounting Medium: Catalog No. 1111.
Ready-to-use dropper vial containing 5.0 ml glycerol-
based mounting medium, pH 9.1 ± 0.2. This reagent
contains 0.1% sodium azide as a preservative.
Coverslips: Catalog No. 1041. Each packet contains
ten 24x60 mm No. 1 glass coverslips.
ADDITIONAL MATERIALS REQUIRED
PROVIDED)
Volumetric pipettes to deliver 20-25 µl volumes
Three Coplin jars or staining dishes
Squeeze bottle or Pasteur pipettes
Serological pipettes
One-liter screw cap containers (for PBS buffer)
Closed container for storing Colorzyme® color
reagent.
Deionized or distilled water
Test tubes to prepare serum dilutions
Bibulous paper or paper towels
Chamber for incubation
Disposable latex gloves
Lab timer
Standard light microscope capable of 200X and 400X
magnification
PRECAUTIONS
1. All human source materials used for this product
have been tested and found to be negative (not
repeatedly reactive) for antibodies to Human Im-
munodeficiency Virus-1 (HIV-1), Human Immuno-
deficiency Virus-2 (HIV-2), hepatitis C virus (HCV),
and for hepatitis B surface antigen (HBsAg) by FDA
approved methods. However, no test method can
offer complete assurance that HIV-1, HIV-2, hepa-
titis C, hepatitis B, or other infectious agents are
absent. Thus, all kit materials should be handled in
the same manner as potentially infectious materials.
2. All patient samples should be handled at the Bio-
safety Level 2 as recommended for any potentially
infectious human serum or blood specimen in the
Centers for Disease Control/National Institutes of
Health Manual: Biosafety in Microbiological and
Biomedical Laboratories, 1984 Edition.
3. Dilution of the components or substitution of com-
ponents other than those provided in this system
may yield inconsistent results.
4. Sodium azide (0.1%) is used as a preservative. So-
dium azide may react with lead or copper plumb-
ing and form explosive metal azide salts. When
disposing of reagents, flush with ample volumes of
tap water to prevent potential residues in plumb-
ing. Sodium azide is a poison and may be toxic if
ingested.
5. This kit is for in vitro diagnostic use.
6. In the event hemolyzed or lipemic sera must be
used, heat inactivate sera 30 minutes at 56°C for
optimal results. Microbially contaminated sera
should not be used.
7. It has been demonstrated that false positive ANA's
may occur due to C1q binding to DNA, indepen-
dent of the substrate used (30). To eliminate poten-
tial C1q interference, routinely heat inactivate all
patient sera at 56°C for 30 minutes prior to testing.
8. The titratable control serum is intended for use in
monitoring lot-to-lot and run-to-run reproducibility.
It is not intended as a measurement of overall
sensitivity or specificity of the assay.
9. Do not smoke, eat, or drink in areas where speci-
mens or kit reagents are handled.
10. Avoid splashing or generation of aerosols at all
11. Incubation times and temperatures other than
12. Cross contamination of reagents or samples may
13. Reusable glassware must be washed and thor-
14. Bring all reagents, slides, and specimens to room
15. Wear disposable latex gloves when handling speci-
16. Microbial contamination of reagents or samples
17. Never pipette by mouth and avoid contact of
18. Color reagent may be reused for up to 30 days
SPECIMEN COLLECTION
Collection: Serum is the preferred specimen. Approxi-
mately 5 ml of whole blood should be collected asepti-
cally by venipuncture using a sterile vacuum collection
(BUT NOT
tube or other suitable collection system. Allow blood to
clot at room temperature (18-24°C). Serum should be
separated from the clot by centrifugation as soon as
possible to minimize hemolysis.
Interfering Substances: Sera exhibiting a high degree
of hemolysis, icterus, lipemia, or microbial growth
should not be used because these conditions may
cause aberrant results. Specimens containing visible
particulate matter should be clarified by centrifugation
before testing.
Storage: Sera may be stored at 2-10°C up to one week.
If testing is further delayed, sera should be stored frozen
at -20°C or lower. Serum should not be stored in a self-
defrosting refrigerator or freezer.
CAUTION: Repeated freeze/thawing of patient samples
may yield false positive or false negative results.
INTERPRETATION OF RESULTS
QUALITY CONTROL
Positive, negative and PBS controls should be run in the
wells provided for quality control on each slide. The
positive control should show dark blue-purple staining of
the nuclei of the cells, with a clearly discernible pattern
characteristic of the control serum that was used. The
cytoplasm may stain light blue-purple in the positive
control well. The negative control should show light
blue-purple staining of both the cytoplasm and nucleus,
but with no clearly discernible pattern of nuclear stain-
ing. The PBS control is used to observe non-specific
staining of the enzyme antibody reagent, and should
not exhibit blue staining. If the controls do not appear
as described, the test is invalid and should be repeated.
OPTIONAL TITRATABLE CONTROL
When reading titers, many laboratories begin reading
with the well that contains the most dilute sample and
read "backwards" to the 1:40 dilution. The first well in
which a clearly discernible nuclear staining pattern is
visible is the titer end-point. We recommend this tech-
nique for determining titer end-points.
The mean titer and titer range (± one dilution on either
side of the mean) determined were established in our
laboratory and are stated as a guide. This control is
provided to allow each laboratory to assess the repro-
ducibility (precision) of its ANA testing. Since this control
is not intended to be an indicator of titer accuracy,
each laboratory should establish its own mean titer end-
point for this sample, and should use this information to
assess run-to-run reproducibility (precision).
Through multiple testing of this titratable control, using
the Immuno Concepts Colorzyme® ANA Test System,
a mean titer value has been established for each lot
number. The lot number, mean titer and titer range (±
one twofold dilution on either side of the mean) are
stated on the vial label and should be used as a guide
for the test system performance.
It is important that the intensity of staining not be
confused with the presence or absence of antinuclear
4
times.
those specified may give erroneous results.
give false results.
oughly rinsed free of detergents prior to use. All
glassware must be clean and dry before use.
temperature (18-24°C) prior to use.
mens and reagents, and wash hands thoroughly
afterwards.
may give false results.
reagents and specimens with skin and mucous
membranes. If contact occurs, wash with a germi-
cidal soap and copious amounts of water.
or until any color change or precipitate is visible.
Cloudiness or opalescence, with no visible precipi-
tate upon reuse is normal. Depending on the rate
of use, 150 ml of Colorzyme® color reagent can be
used with up to twenty slides.