WELL BEFORE USING.

RF 2. RF Positive Control Serum: A stabilized, prediluted human serum

Latex Test Kit 3.
containing at least 8 IU/mL of RF.
RF Negative Control Serum: A stabilized, prediluted human serum
containing less than 8 IU/mL of RF.
320-100 4. Glycine-Saline Buffer (20x): pH 8.2 ± 0.1M glycine and 0.15M NaCl
5. Reaction Slide.
6. PipetteStir Sticks.

NOTE:
1. Dilute buffer following instructions on the label before using.
2. All reagents contain 0.1% (wv) sodium azide as a preservative.
100 Tests 3. The human sera used in the controls has been tested and found negative for
HbsAg and HIV, however, careful handling is always recommended.
4. Store all reagents at 2 - 8°C. DO NOT FREEZE.
INTENDED USE
Materials required but not provided
For the qualitative and semi-quantitative measurement of RF in human serum.
1. Timer.
2. Test Tubes.
SUMMARY 3. Serological pipettes.

Rheumatoid factors (RF) are antibodies directed against antigenic sites in the
Fc fragment of human and animal IgG. Their frequent occurrence in PRECAUTIONS
rheumatoid arthritis makes them useful for diagnosis and monitoring of the
disease. 1,2 1. Reagents containing sodium azide may combine with copper and lead
plumbing to form highly explosive metal azides. Dispose of reagents by
One method used for rheumatoid factor detection is based on the ability of flushing with large amounts of water to prevent azide buildup.
rheumatoid arthritis sera to agglutinate sensitized sheep red cells, as observed 2. For In Vitro diagnostic use. Positive and negative controls prepared using
by Waaler3 and Rose. 4 A more sensitive reagent consisting of biologically human sera were found negative for hepatitis B surface antigen (HBsAg)
inert latex beads coated with human gamma globulin was later described by as required by FDA; however, handle controls as if potentially infectious.
Singer and Plotz. 5 The RF kit is based on the principle of latex agglutination
assay by Singer and Plotz. 5 The major advantage of this method is rapid
performance (3 minute reaction time) and lack of heterophile antibody ASSAY PROCEDURE
interference.
Qualitative Test:
1. Bring reagents and specimens to room temperature before use.
TEST PRINCIPLE 2. Place one drop (50 µl) of the RF Positive Control on field #l of the reaction
slide. Place one drop (50 µl) of the RF Negative Control on field #2. The
The RF reagent is a suspension of polystyrene latex particles sensitized with remaining fields are used for test specimens.
specially prepared human IgG. The reagent is based on an immunological Using pipettes provided. Place one drop of the undiluted specimens on
reaction between human IgG bound to biologically inert latex particles and successive fields. Retain PipetteStir Sticks for mixing step.
rheumatoid factors in the test specimen. When serum containing rheumatoid 3. Gently resuspend the RF Latex Reagent and add one drop to each test field.
factors is mixed with the latex reagent, visible agglutination occurs. The RF Use pipetteStir Stick to spread reaction mixture over entire test field.
latex reagent sensitivity has been adjusted to detect a minimum of 8 IU/mL of 4. Rotate the slide manually or with a mechanical rotor at 80-100 rpm for 2
rheumatoid factors according with the WHO International Standard without minutes and read immediately under direct light.
previous sample dilution. 5. Presence of agglutination of the latex particle is a positive result (see figure
1). Agglutination indicates a RF concentration of equal or more than 8
SPECIMEN COLLECTION AND PREPARATION IU/ml. Sera with positive agglutination should be run again with the
Quantitative Test.
1. Use fresh serum collected by centrifuging clotted blood.
2. If the test cannot be carried out on the same day, the serum may be stored Quantitative Test:
between 2 - 8°C for no longer than 72 hours after collection. 1. Bring reagents and specimens to room temperature before use.
3. For longer periods the sample must be frozen. 2. Using the Glycine-Saline Buffer, dilute the specimens 1:2, 1:4, 1:8, 1:16,
4. As in all serological tests, hemolytic or contaminated serum must not be 1:32 or as needed.
used. 3. Place one drop (50 µl) each of negative and positive controls on two slide
5. Do not use plasma. rings. Place on drop (50 µl) of each dilution on successive fields of the
reaction slide.
4. Gently resuspend the RF Latex Reagent and add one drop to the reaction
MATERIALS AND COMPONENTS slide.
5. Gently resuspend the RF Latex Reagent and add one drop to each test field.
Use Pipette/Stir Stick to spread reaction mixture over entire field.
Materials provided with the test kits 6. Rotate the slide for 2 minutes and read immediately under direct light.
1. RF Latex Reagent: A suspension of uniform polystyrene particles coated
with IgG (human) in glycine buffer, pH 8.2; reagent sensitivity is
standardized with the World Health Organization RF Standard. MIX

COR CODE #26 Page 1 of 2

QUALITY CONTROL EXPECTED VALUES
1. RF Positive and Negative Control should be included in each test batch. 1. The diagnosis of rheumatoid arthritis is based largely on clinical
2. Acceptable performance is indicated when a uniform milky suspension examination, but laboratory tests are useful to support the clinical
with no agglutination is observed with the RF Negative Control and diagnosis and to evaluate the severity and course or the disease in the
agglutination with large aggregates is observed with the RF Positive individual patient. One of the most useful clinical markers for rheumatoid
Control. (Figure 1) arthritis is rheumatoid factor in serum. Rheumatoid factor is a term used
to describe a variety of antibodies or immune complexes or both, that
occur with rheumatoid arthritis as well as in a variety of other diseases. 8
RESULTS 2. Different studies have shown positive serological reactions for rheumatoid
factor in as high as 90 of patients with rheumatoid arthritis compared
Qualitative Test: with less than 5 in control groups. 1
 Negative Result: A negative reaction is indicated by a uniform milky
suspension with no agglutination observed with the RF Negative Control.
 Positive Result: A positive reaction is indicated by any observable
agglutination in the reaction mixture. The specimen reaction should be STORAGE
compared to the RF Negative and Positive Controls (Fig. 1).
1. Reagents are stable until stated expiration date on bottle label when stored
Semi-quantitative Test:The titer of the serum is the reciprocal of the highest refrigerated (2 - 8°C).
dilution, which exhibits a positive reaction. An estimate of the RF 2. DO NOT FREEZE.
3. The RF latex reagent, once shaken must be uniform without visible
concentration in the specimen can be expressed in IUml by using the
clumping. When stored refrigerated, a slight sedimentation may occur and
following equation:
should be considered normal.
4. Do not use the latex reagent or controls if they become contaminated.
IUml of specimen = IUml control  specimen titer

REFERENCES
1. Taborn. J.D., et al. Rheumatoid Factor: 1 Review, Lab. Med. 0:392 1979).
2. Dornerm R.W., et al. "Critical Review Rheumatoid Factor".Clin. Chem.
Acta 167:1 (1987).
3. Waaler. E., Acta. Path. Microbial Scan. 17:172 (1942).
4. Rose. H.M., et al., Proc. Soc. Exp. Biol. Med 68:1 (1943).
5. Singer. J.M., et al., .Am. J. Med 21:888 (1956).
6. Winchester. R. Am. Soc. for Micro 665 (1976).
7. Rothermich. N.O., et al. J.A.M.A. 164:1999 (1957).
Positive Negative 8. Hansen. S.L., et al. Am. J. Clin. Pathol. 73:110 (1980).
Figure 1.

PERFORMANCE CHARACTERISTICS ISO 13485

ISO 9001
1. Sensitivity: 8 IUml or above.
2. Comparison:
A. Qualitative Results: The RF Latex Reagent was evaluated on a total
of 75 samples from the hospital patients. The qualitative test was
evaluated by comparison with a commercially available latex
agglutination test. This study demonstrated a 96 agreement Diagnostic Automation/Cortez Diagnostics, Inc.
between these tests. The discrepancy results were obtained in 21250 Califa Street, Suite 102 and 116,
samples with titers near the limit of sensitivity of the reagents. Woodland Hills, California 91367 USA
B. Semi-quantitative Results: A panel of 10 known RF positive serum.
samples was quantitated on three consecutive days. The results of
Date Adopted 2017-03-16
the study indicated that RF Latex Reagent has 100% precision.
CORTEZ- RF
320-100
Latex Test Kit-2016
CEpartner4U , Esdoornlaan 13,
LIMITATIONS OF PROCEDURE 3951DB Maarn. The Netherlands.
1. Results should be read two (2) minutes after the mixing or the reagent on www.cepartner4u.eu
the slide. Revision Date: 2016-Aug
2. Existence of prozone at high titers has not been encountered.
3. Increased levels of RF may be round in some diseases other than
rheumatoid arthritis such as infectious mononucleosis, sarcodosis, lupus
erythematosus, Sjogren's syndrome. 6,7
4. Certain patients with rheumatoid arthritis will not have the RF present in
their serum. 6

COR CODE #26 Page 2 of 2