1. health and fitness
2. disease

golgi, ribosmomes, lysosomes, mitochondria and cytoskeletal elements (smooth muscle
antibodies) should be followed up with appropriate frozen tissue sections. HEp-2 substrates are
not equally sensitive for mitochondrial and smooth muscle testing when compared with tissue
section antigens.
ANTINUCLEAR ANTIBODY TEST SYSTEM
HEp-2
REF 10-1120
120 Tests
Store kit at +2 to +8°C
Pour d'autres langues
Für andere Sprachen
Para otras lenguas
Per le altre lingue
Dla innych języków
Para outras línguas
Για τις άλλες λώσσες
För andra språk
For andre språk
wwww.trinitybiotech.com
INTENDED USE
MarDx Antinuclear Antibody Test System is intended for testing human serum for the presence of
human antinuclear antibodies (ANA) as an aid in the diagnosis of chronic autoimmune disorders.
For in vitro diagnostic use. High complexity test.
SUMMARY AND PRINCIPLES
Indirect fluorescent antibody (IFA) Techniques have been used extensively for the detection of
antinuclear antibodies (ANA) in patient sera for diagnostic evidence, prognostic significance and
therapeutic management. Screening for these antibodies is routinely done on patients with
various connective tissue diseases, particularly in Systemic Lupus Erythematosis (SLE) which
may be autoimmune or drug induced (see Table I). Drug induced lupus is differentiated from
classic lupus by the absence of anti- DNA antibodies and the presence of anti-histone antibodies.
TABLE I
SLE INDUCING DRUGS
Group I:
Inducted by Pharmacological Action
Hydralazine
Procainamide
Anti-convulsants:
Mephenytoin
Phenytoin
Trimethadione
Ethosuximide
Carbamazepine
Pheneturide
Isoniazid
Chlorpromazine
1. HOMOGENEOUS PATTERN:
Fluorescence is uniform and diffuse in the nucleus of the interphasic cells.
Metaphasic cells demonstrate homogeneous fluorescence of the condensed chromosomal
region
Specificity: The homogeneous pattern is obtained with antibodies to DNA, DNP, and histones.
2. PERIPHERAL PATTERN:
Fluorescence is uniform and diffuse with staining of a greater intensity at the outer region of
the nucleus.
Metaphasic cells demonstrate strong staining of the condensed chromosomal submembranal
region.
Specificity: The peripheral pattern is obtained with antibodies to DNA and histone.
3. COARSE SPECKLED WITHOUT NUCLEOLUS PATTERN:
Fluorescence is uniform with coarse specks in the nuclear matrix with no nucleolar staining.
Metaphasic cells demonstrate no staining of the condensed chromosomal region.
Specificity: The coarse speckled pattern without nucleolus staining is obtained with anti- SM
and anti-RNP antibodies.
4. COARSE SPECKLED WITH OCCASIONAL NUCLEOUS PATTERN:
Fluorescence is uniform with coarse flat specks in the nuclear matrix with occasional
nucleolar staining.
Metaphasic cells demonstrate no staining of the condensed chromosomal region.
Specificity: The coarse speckled pattern with occasional nucleolar staining is obtained with
anti-SS-B antibodies.
5. FINE SPECKLED WITHOUT NUCLEOLUS PATTERN:
Fluorescence is uniform with fine specks in the nuclear matrix without nucleolar staining.
Metaphasic cells demonstrate no staining of the condensed chromosomal region.
Specificity: The fine speckled pattern without nucleolus staining can be found with several
antibodies including SS-A, Mi-1, Mi-2, and SL.
Group II:
Induced by Allergic Action
Aminosalysilic Acid
Chlorthalidone
D-Penicillamine
Griseofulvin
Isoquinazepon
Levodope
Methyldopa
Methysergide
Methylthiouracil
Oral Contraceptives
Oxyphenisatin
TABLE II
CLASSIFICATION OF CLINICALLY SIGNIFICANT SINGLE PATTERN ANA REACTIONS
USING HEP-2 CELLS.
Pencilillin
Phenyibutazone
Practolol
Propylthiouracil
Quinidine
Reserpine
Streptomycin
Sulfonamides
Tetracycline
Tolazamide
The detection of positive antinuclear antibodies depends in part on the antigenic substrate and
the fixation method used in the preparation of the substrate. ANA testing can utilize many
substrate sources such as liver or kidney tissue sections which can be derived from rats or mice.
The use of human tissue culture cells (specifically HEp-2) has provided an alternative substrate
to tissue sections.
HEp-2 substrate allows for differentiation of pattern recognition on cells in all phases of mitosis
which is not necessarily possible using tissue sections as substrate. The high level of mitotic
cells in tissue culture HEp-2 cells allows for the definitive differentiation of "centromere" antigen
staining of the condensed chromosomal region of metaphasic cells. Centromere positive sera
are generally reported as "speckled" staining patterns when tissue sections are used.
The high level of mitotic cells found in HEp-2 cells also aids in the determination of mixed
homogeneous and speckled antigenic patterns. Most speckled patterns stain the nucleoplasm
surrounding the mitotic cell but do not stain the condensed chromosomal region of the
metaphasic HEp-2 cells. Homogeneous patterns will stain the condensed chromosomal region
with an intense homogeneous or peripheral stain. Therefore, staining in the same mitotic HEp-2
cell of both homogeneous condensed chromatin of metaphasic cells and a speckled surrounding
nucleoplasm of metaphasic cells indicates the presence of two types of nuclear specificities in
the same sample. Mixed homogeneous/speckled specificities using tissue sections are generally
observed upon titration of the sample which reveals patterns present in conjunction with the
homogeneous pattern.
SS-A antigens are specifically fixed in the HEp-2 substrate preparation to assure its reproducible
abundance in testing for this clinically important antigenic determinant. Tissue sections may vary
in the amount of SS-A present from manufacturer to manufacturer. Anti-SS-A is found in
neonatal lupus syndrome, Sjogren's Syndrome and SLE. Anti-SS-A antibodies bind to skin and
heart tissue. Study of expectant mothers with rheumatic disease symptoms for the presence of
anti-SS-A is recommended to prevent untoward neonatal death.
Postpartum clinical monitoring of mothers of children with heart block and rashes for the
development of clinical and serologic SLE is clinically indicated. Both ANA testing with
immunofluorescence and double gel diffusion testing with extractable nuclear antigens (ENA) are
recommended for detecting and typing the Anti-SS-A response. Proliferating cell nuclear antigen
is also detected in HEp-2 cells which may vary in tissue section substrates. Other nuclear
patterns may be observed in HEp-2 cells which are not yet clinically defined. They include
staining of the mitotic cell spindle apparatus including mitotic spindle fibers, centriole,
chromosomes, midbody of telophase cells and nuclear cell membrane. Additionally, non-nuclear
cytoplasmic organelles can stain with various staining patterns. Generally cytoplasmic staining of
6. FINE SPECKLED WITH NUCLEOLUS PATTERN:
Fluorescence is uniform with fine specks in the nuclear matrix with nucleolar staining.
Metaphasic cells demonstrate staining of the condensed chromosomal region.
Specificity: The fine speckled with nucleolus pattern is demonstrated by antibodies to Scl-70.
This pattern is considered a marker antibody specific for proteinkinase (70K) and is
clinically diagnostic of scleroderma.
7. DIFFUSE SPECKLED (CENTROMERE) WITHOUT NUCLEOLUS PATTERN:
Fluorescence is uniform with spherical specks approximately 40-60 per cell nucleus.
Metaphasic cells demonstrate intense speckled staining of the condensed chromosomal
region corresponding to the centromere region.
Telophase cells do not demonstrate speckles.
Specificity: Diffuse speckled with centromeric staining of metaphasic cells is demonstrated by
antibodies to the kinetocore antigens of the centromere. This pattern is considered a
marker antibody and is diagnostically specific for the CREST syndrome.
8. ATYPICAL DISCRETE SPECKLED (PSEUDOCENTROMERE, NUCLEAR DOTS):
Occasional speckled fluorescence of less than 10 bright dots in the nuclei of interphasic cells
only.
Metaphasic cells demonstrate no speckling of condensed chromosomal region.
Specificity: The chemical nature of the pseudocentromeric pattern is not yet known. The
antibody has been defined as an anti Nsp-1. This antibody is associated with PBC and
Chronic Hepatic Disease.
9. PLEOMORPHIC SPECKLED OF VARIOUS INTENSITIES:
Varying fluorescence of fine to coarse speckling in approximately 30-60% of the cells. Both
the number of cells staining and the intensity of the speckles vary.
Metaphasic cells may demonstrate positive or negative staining of the condensed
chromosomal regions.
Specificity: The antibody responsible for pleomorphic speckled fluorescence is proliferating
cell nuclear antibody (PCNA) and is active against cyclin.
10. NUCLEOLAR ONLY:
The staining of the nucleolus may be either homogeneous, clumpy, speckled or perinuclear.
Metaphasic cells generally do not show staining of the condensed chromosomal regions.
Positive staining can be seen in the nucleolus of cells at the end of anaphase in telophase.
Specificity: The antibodies responsible for staining of the nucleolus include: Anti-4-6 RNA,
Anti-RNA and RNP, and Anti-DNA Nucleolar Organizer.
Page 1 of 4-EN
10-1120-29-Rev. 2
SUMMARY AND PRINCIPLES (continued)
CYTOPLASMIC ORGANELLE PATTERNS:
1. FINE CYTOPLASMIC:
(Jo-1) Fine perinuclear cytoplasmic fluorescence.
Specificity: The antibodies responsible for staining of the perinuclear bodies are designated
as Jo-1 and are active against a histidyl-modified t-RNA synthetase moiety. The Jo-1
antibody is considered a diagnostic marker antibody for polymyositis where it has a 31%
disease association. The use of gel diffusion tests to identify this antigen is recommended,
as its characterization from other HEp-2 cytoplamic patterns is difficult.
2. COARSE CYTOPLAMIC:
(Anti-mitochondrial) Coarse cytoplasmic fluorescence extending around the nucleous along
the cytoskeletal filaments. (anti-mitochondrial)
Specificity: The antibodies responsible for mitochondrial fluorescence include antibodies
against various mitochondrial specificites including M1, M2, M3, M5, and M6. HEp-2 cells
detect the M2 type antigen. However, the use of kidney tissue sections is recommended
since the kidney section has greater sensitivity in the detection of all mitochondrial antibody
specificities for mitochondrial antibody testing.
Nonspecific patterns resembling
mitochondrial reactions on HEp-2 cells may occur and nonspecific mitochondrial-like HEp-2
patterns will be negative using tissue sections. Therefore, HEp-2 mitochondrial patterns
should be confirmed on tissue sections.
3. FILAMENTOUS OR FIBROUS CYTOPLASMIC:
(Anti-smooth muscle) Cytoskeletal fluorescence surrounding the nucleus. The fluorescence
may occasionally appear as fine perinuclear radial filaments with coils due to vimentin
antibodies or they may occasionally appear as fine fibrous staining seen with antibodies to
actin. Smooth muscle staining of tissue sections due to desmin will not stain the
cytoskeleton of HEp-2 cells.
Specificity: As with mitochondrial antibody testing tissue section substrates are more specific
and sensitive than HEp-2 cells for routine testing of all three types of anti-smooth muscle
antibodies.
The primary reaction occurs during the first incubation period while the patient's serum covers
the substrate.
The secondary reaction follows a PBS rinsing to remove any unbound human antibody. The
reagent used in the secondary reaction is a fluorescein labeled anti-human conjugate which has
been affinity purified for use with Hep-2 cell culture substrates and has been adjusted for the
optimum use dilution and is free of most nonspecific staining of the HEp- 2 cell.
After a second PBS rinse to remove any unbound anti-human globulin conjugate, the specific
type of nuclear fluorescence observed and its fluorescence intensity is reported visually using a
fluorescence microscope at a magnification of 400X. (See Table II).
1.
2.
3.
4.
5.
6.
7.
The disease association of various ANA antibodies in Systemic Rheumatic Diseases is
summarized in Table III.
8.
9.
TABLE III
ASSOCIATION OF ANA WITH SRD
ANTIBODIES TO:
DS-DNA only
DA and ssDNA
SS-DNA only (exposed purine and
pyrimidines)
Histones
Non-Histone Antigens
Sma
nuclear RNP (u! RNP)
SS-B/La
PCNA
Ma
Scl-70a
Centromere/kinetochorea
RANA
PM-1 (PM/Sol)
Mi-1
J0-1a
Ku
Nucleoli
4-6s RNA
RNA and RNP
DNA:nucleolar organizer
DISEASE ASSOCIATION
SLE:rare cases
SLE:60-70%, in other diseases may occur
in low titer
SLE:other rheumatic diseases, some nonrheumatic diseases
SLE:60-70%
Drug LE (procainamide): 95%
Other drug LE: 30-90% RA:30% low titer
10.
11.
12.
13.
PRECAUTIONS
Always wear suitable protective clothing, gloves and eye/face protection when working with
this product.
Each donor unit used in the preparation of this material was tested by an FDA approved
method for the presence of the antibody to HIV as well as for HBsAg and found to be
negative (were not repeatedly reactive). WARNING - POTENTIAL BIOHAZARDOUS
MATERIAL Because no test method can offer complete assurance that human
immunodeficiency virus (HIV), hepatitis B virus, or other infectious agents are absent, these
human control reagents should be handled at the Biosafety 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", 1999. (23)
The phosphate buffered saline and mounting medium found in this kit are irritating to the
eyes, respiratory system and skin.
Some components in this kit contain 0.1% Proclin 300. At full strength Proclin 300 is
corrosive and will cause burns and possibly sensitisation by skin contact.
The conjugate in this kit contains 0.015% Evan’s Blue. Evan’s Blue is a possible
carcinogen and may cause reproductive harm.
Some components in this kit contain 0.02% Thimerosal. Thimerosal is toxic by inhalation, in
contact with skin, and if swallowed, and is a reproductive hazard.
The conjugate and controls in this kit contain sodium azide at a concentration of less than
0.1 %. Sodium azide is toxic if ingested and forms potentially explosive copper and lead
azide compounds in waste plumbing lines. Should the reagents come in contact with copper
or lead waste plumbing, flush the waste line with large quantities of water to prevent the
formation of potentially explosive compounds.
Do not use components beyond their expiration date.
Follow the procedural instructions exactly as they appear in this insert to insure valid
results.
For in vitro diagnostic use.
Handle slides by the edges since direct pressure on the antigen wells may damage the
antigen.
All reagents must be brought to 20 to 25°C before performing the test procedure.
Once the procedure has been started do not allow antigen in the wells to dry out. This may
result in false negative test results, or unnecessary artifacts.
R43:
S28-37:
SLE:25-40%, marker antibody
MCTD: 95-100%,
lower frequency in SLE, discoid LE, PSS
SS-A/Ro
Sjogren's Syndrome: 60-70%
SLE: 30-40%
Neonatal Lupus Syndrome: 100%
Sjogren's Syndrome: 50-60%
SLE:10-20%
SLE:<10%
SLE:20%
SS: 15-20%
Prod#
10-1012
10-1202
10-1206
10-1201
10-1513
90-1612
90-1607
90-1700
90-1712
CREST syndrome: 70-90%
RA: 90-95%
Polymyositis/scleroderma overlap: 64%
Dermatomyositis: 17%
Dermatomyositis: 11%
Polymyositis: 31%
Polymyositis/SS overlap: (55%)
SS and certain overlap diseases
Sjogren's Syndrome
Sjogren's Syndrome
1.
2.
3.
4.
5.
6.
1.
2.
a Marker Antibody = Presence of antibody is virtually diagnostic.
The HEp-2 substrate utilized in this IFA test system contains a mixture of cells in various mitotic
stages. The majority of the cells are in a resting phase (interphase), the ideal phase for routine
ANA screening. The remainder are a mixture of the four phases of mitosis: 1. prophase 2.
metaphase 3. anaphase 4. telophase. When interpreting the patient sample, both resting and
mitotic cells should be evaluated.
ANA antibodies are not organ or species specific. The primary test reaction involves circulating
antinuclear antibodies present in the patient's serum which attach to their homologous nuclear
antigens.
3.
4.
5.
6.
Xi - Irritant
May cause sensitization by skin contact
After contact with skin, wash immediately with plenty of water and soap. Wear
suitable gloves
MATERIALS PROVIDED
Description
HEp-2, 12 well slide
ANA (4+) Homogeneous Positive Control
ANA (1+) Homogeneous Positive Control
Autoimmune Negative Control
FITC Conj., HEp-2 with Evans’ Blue (Goat)
FITC HEp-2 Mounting Medium (pH 7.5)
Phosphate Buffered Saline (pH 7.5)
Coverslips, 70x22 mm
Blotters, 12 well
Quantity
10 ea
0.5mL
0.5mL
0.5mL
7.0mL
3.0mL
2x10g
12 ea
10 ea
ADDITIONAL MATERIALS REQUIRED BUT NOT PROVIDED
Test tubes, test tube rack, pipettes.
Volumetric flask (1 liter)
Staining dish.
Epifluorescence microscope
Microscope Slide Roller
Humid Chamber
STORAGE AND STABILITY
Antigen slides Prod# 10-1012 should be stored at +2 to +8°C. Slides are stable until their
expiration date on the product label.
Positive control Prod# 10-1202 and Prod# 10-1206 should be stored at +2 to +8°C. Refer to
expiration date on label.
Negative control Prod# 10-1201 should be stored at +2 to +8°C. Refer to expiration date on
label.
FITC labeled anti-human conjugate Prod# 10-1513 should be stored at +2 to +8°C. Refer to
expiration date on label.
Mounting Medium Prod# 90-1612 should be stored at +2 to +8°C. Refer to the expiration
date on label.
Phosphate buffered saline pH 7.5 Prod# 90-1607 are stable at room temperature.
Reconstitute each vial of PBS buffer salts with 1.0L of distilled water. The PBS contains no
preservative and should be stored at +2 to +8°C. Discard if turbidity develops.
Page 2 of 4-EN
10-1120-29-Rev. 2
TEST PROCEDURE
Dilute test serums 1:40 in PBS if testing is being performed for screening purposes. For titrations
set up doubling dilutions of serum starting at 1:40, (i.e. 1:40, 1:80, 1:160, 1:320, etc.). The slide,
controls and conjugate are ready to use.
1.
Tear envelope at notch. Carefully remove the slide and avoid touching the antigen areas.
The slide is now ready to use.
2.
Place a drop of diluted serum (15 to 20 l) over the antigen wells.
3.
Place slide with patient's serum and controls in a moist chamber for 30 minutes at room
temperature. (approximately 20°C)
4.
Remove slide from moisture chamber. Using a wash bottle, gently rinse remaining sera
from slides being careful not to aim the stream directly on the well.
5.
Wash in PBS for two separate five minute changes
6.
Remove the slides from PBS and place slide antigen side facing up on a dry paper towel.
Carefully place the blotter over the slide so that the blotter is indexed to the surface of the
microscope slide. Hold one edge of the blotter with one hand to keep the blotter in place
and apply sufficient gentle pressure with the microscope slide roller to remove the
moisture between the antigen wells. DO NOT ALLOW THE ANTIGEN WELLS TO DRY.
7.
Using dispenser provided, deliver 1 drop (25 l) of conjugate per antigen well. Repeat
steps 3-6.
8.
Place 4-5 drops of mounting medium on slide.
9.
Apply a 22 x 70mm cover glass. Examine the slide under a fluorescent microscope. Note:
To maintain fluorescence, store mounted slide in a humid chamber placed in a dark
refrigerator.
*
The conjugate dispenser is provided with a calibrated tip and allows quantitative delivery
of reagents from the storage battle. To use, wipe the tip with a paper towel, invert the
bottle and squeeze gently to release one drop. If the tip contains an air bubble, tap the
battle gently to remove air bubble which will ensure precise drop delivery.
SPECIMEN COLLECTION AND STORAGE
Serological specimens should be collected under aseptic conditions. Hemolysis is avoided
through prompt separation of the serum from the clot. Serum should be stored at 2°C to 8°C if it
is to be analyzed within 4-7 days. Serum may be held for 3 to 6 months by storage at -20°C or
lower. Lipemic and strongly hemolytic serum should be avoided. When specimens are shipped at
ambient temperatures, additions of a preservative such as 0.01% thimerosal (merthiolate) or
0.1% sodium azide is strongly recommended. The CLSI provides recommendations for storing
blood specimens (Approved Standard Procedure for the Handling and Processing of Blood
Specimens, H18-A2 2005). (24)
TITER INTERPRETATION
The titer is the highest dilution of patient's serum showing weak (1+) fluorescence. Less than
1:40 Normal; virtually rules out active SLE provided patient is not on immunosupressive therapy
or in remission.
1:40-1:80
1:160 or greater
Positive; test often in RA and other connective tissue diseases.
A fresh sample should be tested in two weeks. If the titer increases
active SLE is suggested. No change in titer indicates other possible
autoimmune disease in a static condition or a treated controlled SLE
patient. A decrease in titer indicates an SLE case in remission or a
treated controlled SLE case or another autoimmune process.
Strongly suggests SLE although other autoimmune diseases and
drugs may induce these high titers.
PATTERN INTERPRETATION
ANA patterns are generally reported as: Homogeneous, Peripheral, Speckled, Centromere,
Nucleolar and in multiple combinations. The nuclear ANA patterns found in IFA may be of
diagnostic and/or prognostic significance. (SEE TABLE II & TABLE III).
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
LIMITATIONS OF PROCEDURE
No diagnosis should be based upon a single ANA test result, since various host factors
must be taken into consideration.
Among these host factors are sex and age. There is an increasing incidence in positive
ANA results in both males females as age increases (10). Normal females between 20-60
have 7% incidence of ANA: normal males, a 3% incidence. Normal males and females
over 80 years of age have a 50% incidence of ANA.
Various medications including antibiotics, tranquilizers, aspirin and birth control pills can
induce a Lupus-Iike condition resulting in high ANA titers (11). (See table I) Drug- induced
Lupus generally goes into sustained clinical remission following removal of the triggering
medication.
Various autoimmune processes induce positive ANA tests.
Further evidence for a diagnosis of SLE is provided by low complement levels, particularly
C1Q, C3 and C4. (12).
ANA tests may not agree with LE Prep tests or with latex tests.
Management of therapy should be based not only on positive serologic tests for SLE, but
should include the presence of active clinical disease. (13).
Elderly patients with SLE have a better prognosis and their clinical symptoms differ
substantially from those seen in younger patients. (14).
Although the predominant class of antinuclear antibodies (ANA) is immunoglobulin G, the
presence of immunoglobulin E maybe of pathogenic importance in SLE (15).
Staining patterns often change with titration of the sera revealing multiple patterns not
seen in the lower dilutions.
Identification of antibodies based only upon patterns could be misleading and should be
confirmed using other serological tests such as ENA double gel diffusion tests, specific
nDNA tests and histone tests.
1.
2.
3.
4.
5.
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
24.
QUALITY CONTROL
Positive 4+, 1+, and negative serum controls must be included in each day's testing to
confirm reproducibility, sensitivity and specificity of the test procedure.
The negative serum control should result in little(+) or no fluorescence of the nuclei. If this
control shows bright fluorescence either the control or the antigen may be at fault.
The positive 4+ serum control should result in 3+ to 4+ fluorescence of the type specified on
the label. If this control shows little or no fluorescence either the control, antigen, conjugate
or technique may be at fault.
The positive 1+ serum control should result in 1+ fluorescence of the type specified on the
label. If this control shows little or no fluorescence either the control, antigen, conjugate or
technique may be at fault.
In addition to positive and negative serum controls, a PBS control should be run to establish
that the conjugate is free from nonspecific staining of the antigen substrate. If the antigen
shows bright fluorescence in the PBS control repeat using fresh conjugate. If the antigen
still fluoresces either the conjugate or the antigen may be at fault.
REFERENCES
Feely RH, Systemic Lupus Erythematosus: A Review. Rheum. and Rehab., 17:79, 1978.
Burnham TK, Antinuclear Antibodies II. The Prognostic Significance of Nuclear
Immunofluorescent Patterns in Lupus Erythematosus. Arch. of Derm., 11.203, 1975.
Greenwald CA, Peebles CI, Nakamura RM, Laboratory Tests for Antinuclear Antibody
(ANA) in Rheumatic Disease. Lab. Med., 9., 1978.
Nisengard RJ, Antinuclear Antibodies: Significance of Titers. Immunology of the Skin by EH
Beutner, TP Chorzelski, SE Bean. 2nd Edition, John Wiley and Sons, p.287. 1979
Barrett EV, Immunofluorescence Tests in Immune Techniques and Applications Amer. Jour.
Clin. Path 68:662, 1977
Lowenstein MB, Rothfield NF, Family Study of Systemic Lupus Erythematosus, Arth. and
Rheum. 20:1293, 1977
Hahon N, Eckert HL, Stewart J, Evaluation of Cellular Substrates for Antinuclear Antibody
Determinations. J. Clin. Microbiol. 2:42, 1975.
Ritchie RH, The Clinical Significance of Titered Antinuclear Antibodies. Arthritis and
Rheumatism. 10.6,1967
Tan EM, Rodnan GP, Garcia I, Moroi Y, Fritzler MJ and Peebles C, Anti Centromere
Antibody and its relationship to Crest Syndrome. Arth and Rheum. 23:6, 1980.
Castanedo JP, White JG, Williams RC Jr. Antinuclear Antibodies in Normal Human
Subjects. Arth and Rheum. 10:5, 1967.
Holborow EJ, Weir DM, Johnson GD. A Serum Factor in Lupus Erythematosus with Affinity
for Tissue Nuclei. British Med Jour Sept 28, 1957.
Clark G, Reichlin M, Tomasi TB Jr. Characterization of a Soluble Cytoplasmic Antigen
Reactive with Sera from Patients with systemic Lupus Erythmatosus. J Immunology 103:1,
1969.
Solomon SD, The PM-1 Antibody Test in a Patient with Rheumatic Complaints. J.Rheum
1982.
Sharp GC, Irvin WS, Tan EM, Gould RG, Holman HR, Mixed Connective Tissue DiseaseAn apparently Distinct Rheumatic Disease Syndrome Associated with a Specific Antibody to
an Extractable Nuclear Antigen (ENA). Am J Med Vol 52, 1972.
Alspaugh M, Maddison, P, Resolution of the Identity of Certain Antigen-Antibody systems in
Systemic Lupus Erythematosus and Sjogren's Syndrome: An Interlaboratory Collaboration,
Frief Reports. p. 796-798. Feb 21, 1979
Cox JV, Schenk EA, Olmstead JB, Human anticentromere Antibodies: Distribution,
Characterization of Antigens, and Effect on Microtubule Organization. Cell Vol 35, p331339, 1983.
Moroi Y, Peebles C, Fritzler M, Steigerwald J, Tan EM. Autoantibody to Centromere
(Kinetochore) in Scleroderma Sera. Proc Natl Acad Sci 77:1627- 1631, 1980
Sluder G, Begg DA, More on the Spatial Arrangement of Spindle Components in the Timing
of Mitotic Events. J Cell Bio Vol 97, 1983.
Nakamura R, Peebles CL, Rubin RL, Molden DP, Tan E, Autoantibodies to Nuclear
Antigens (ANA), Advances in Laboratory Tests and Significance in Systemic Rheumatic
Diseases. ASCP Second Edition, 1985.
McCarthy GA, Autoantibodies and Their Relation to Rheumatic Diseases. Med. Clin. of N.A.
70:237-261 1986.
Fritzler MJ, Tan EM, Antinuclear Antibodies and the Connective Tissue Diseases. Chapter
8. Laboratory Diagnostic Procedures in the Rheumatic Diseases. Grune & Stratton pp207243 1985.
Walravens M, Maini RN, First European ANA Symposium Leuven, Belgium May 1984. CI.
Rheu. Acta Medica Belgica Vol6 Suppl N1, pp 1-108 June 87.
Centers for Disease Control/National Institutes of Health (CDC-NIH) Manual. 1999. In:
Biosafety in Microbiological and Biomedical Laboratories, 4th Edition, U.S. Dept. of Health
and Human Services, Public Health Service.
Clinical Laboratory Standards Institute (CLSI). 2005. Procedures for the Handling and
Processing of Blood Specimens; Approved Guideline – Second Edition. CLSI Publication
H18-A2.
Page 3 of 4-EN
10-1120-29-Rev. 2
Consult Instructions for Use
REF
Product Number
LOT
Lot Number
IVD
In Vitro Diagnostic Medical Device
EC
Authorized Representative in the
European Community
REP
Use By
Caution, consult accompanying
documents
Temperature limitation
Manufacturer
Irritant- Precaution
CONTROL
-
Negative Control
CONTROL
ANA
4+
ANA 4+ Homogeneous Positive Control
CONTROL
ANA
1+
ANA 1+ Homogeneous Positive Control
CONJ
Conjugate
PBS
Phosphate Buffered Saline
MTMED
Mounting Medium
CVRSLP
BLTRS
SLD
Coverslips
12W
12W
EC REP-Trinity Biotech plc.
IDA Business Park
Bray, County Wicklow, Ireland
Phone: +353-1-276-9800
Fax: +353-1-276-9888
Web:www.trinitybiotech.com
Blotters, 12 Well
Slide, 12 Well
Manufactured By
MarDx Diagnostics,Inc.
A Trinity Biotech Company
5919 Farnsworth Court
Carlsbad, CA 92008
Phone: 800-325-3424
Fax: 760-929-0124
10-1120-29-Rev. 2
10/2010
Page 4 of 4-EN
10-1120-29-Rev. 2