Document No.

Page: 1 of 2
Revision Date
Department: Pathology 01 September
2022
Section: Effective Date:
Revision No. 01September 2022

Policy Title:
STANDARD OPERATING PROCEDURE IN RHEUMATOID FACTOR (RF)

I. INTENDED USE

The RA latex test kit is a rapid agglutination procedure for the qualitative
and semi quantitative determination of Rheumatoid factors in human serum
samples.

II. PRINCIPLE

This is a method of detecting rheumatoid factor using a suspension of fine

plastic granules coated with human gamma globulins which were agglutinated in
the presence of rheumatoid factor. The RA latex reagent is sensitive,
standardized preparation of this type, made with a purified human IgG fraction
and selected polysterene latex.

The presence or absence of a visible agglutination indicates the presence

or absence of RF in the sampled tested.

III. CLINICAL SIGNIFICANCE

Rheumatoid factors are found in the sera of most patients with rheumatoid
arthritis as well as variety of other diseases. They are group of antibodies mostly
belonging to the IgM class directed against determinants on the Fc fragments of
patient’s IgG. Rheumatoid factor testing has a high diagnostic value for a tentative
diagnosis based on case history and clinical findings.

IV. REAGENTS AND MATERIALS

1. RED / PLAIN TUBE

2. SEROLOGICAL PIPETTES (50 uL and 100 uL)
3. Buffer solution or NSS
4. Rotator table
5. Timer
6. Glass slides/ Reusable agglutination slides
7. Stirrers
8. Latex reagent

Prepared By: Reviewed By: Noted by: Approved by:

MARIE ANGELICA HABALO, RMT KHRISTINE T. MORENO, RMT CARINA P. VILLAMAYOR,MD,FPSP Roy O. Arceo, MD, MHA, FPSO-HNS
Date: 01 September 2022 Date: 01 September 2022 Date Date:
 Document No. Page: 2 of 2
Revision Date
Department: Pathology 01 September
2022
Section: Effective Date:
Revision No. 01September 2022

Policy Title:
STANDARD OPERATING PROCEDURE IN RHEUMATOID FACTOR (RF)

V. SAMPLE REQUIREMENTS
At least 1mL of freshly collected serum (red/ plain tube)
VI. QUALITY CONTROL
 Control sera should be used daily to verify the test results. International
standards are commercially available.
 Store reagent at 2-8 °C (if not in use)
VII. PROCEDURE
 QUALITATIVE METHOD
1. Allow each component to reach room temperature.
2. Gently shake the latex reagent to dispense the particles.
3. Place a drop of undiluted serum onto a circle of a test slides.
4. Add one drop of the latex reagent next to the drop of serum.
5. Spread the reagent and serum sample over the entire area of the test circle
using a separate stirrer foe each sample.
6. Gently tilt the test slide backward and forward approximately once every two
seconds to two minutes. Positive and negative controls should be included at
regular intervals. Both are ready for use and do not require further dilutions.
 SEMI QUALITATIVE METHOD
The semi- quantitative test can be performed in the same way as the quantitative
test using dilutions of serum in the saline.
DILUTION 1/2 1/4 1/8 1/16
SAMPLE SERUM 100 uL
BUFFER /SALINE 100 uL 100 uL 100uL 100 uL
100 uL 100 uL 100 uL
DIS
POSE
100uL
VOLUME OF 50uL 50uL 50uL 50uL
SAMPLE
200 x No. Dilution 8x2 8x4 8x8 8x16

Prepared By: Reviewed By: Noted by: Approved by:

MARIE ANGELICA HABALO, RMT KHRISTINE T. MORENO, RMT CARINA P. VILLAMAYOR,MD,FPSP Roy O. Arceo, MD, MHA, FPSO-HNS
Date: 01 September 2022 Date: 01 September 2022 Date Date:
 Document No. Page: 3 of 2
Revision Date
Department: Pathology 01 September
2022
Section: Effective Date:
Revision No. 01September 2022

Policy Title:
STANDARD OPERATING PROCEDURE IN RHEUMATOID FACTOR (RF)
I.U. / mL 16 32 64 128
NORMAL LEVEL : <8 I.U. / mL
VIII. CALCULATION
In the semi quantitative test the titer is expressed as the reciprocal of the highest
dilution showing macroscopic agglutination: e.g. if this occurs in dilution 3 the titer is 64.

IX. INTERPRETATION
 Presence of agglutination indicates a level of RA in the sample of equal or
>8mg/L
 No agglutinations indicates a level of RA in the sample of <8mg/L.
 Using a latex system, positive results are not always found with every case of
clinically defined rheumatoid arthritis. The number of positives reported using
various types of latex reagents range from 70 % to over 90% False positive
result also occur in various pathological conditions including lupus
erythematosus, hepatitis, cirrhosis of liver, lymphomas, scleroderma, and
various other infections. The frequency of false positive results is not high even
in these conditions but the possibility must be borne in mind when interpreting
results.

X. LIMITATIONS OF THE PROCEDURE

 The incidence of false positive results is about 3-5%. Individuals suffering from
Infectious mononucleosis, hepatitis, syphilis, as well as elderly people (30%
aged of 60 years +) may give positive results.
 Diagnosis should not be solely based on the result of latex method but also
should be complemented with Waaler Rose test along clinical examination.
 Newborn: results obtained with a latex method do not compare with those
obtained in Waaler Rose test. Differences in the results between methods do
not reflect differences in the ability to detect rheumatoid factors.
 Do not interfere with the with the test results:
 Hemoglobin - 10g/L
 Bilirubin - 20mg/dL
 Lipemia - 10g/L
 Rheumatoid factors > 100IU/mL may interfere

Prepared By: Reviewed By: Noted by: Approved by:

MARIE ANGELICA HABALO, RMT KHRISTINE T. MORENO, RMT CARINA P. VILLAMAYOR,MD,FPSP Roy O. Arceo, MD, MHA, FPSO-HNS
Date: 01 September 2022 Date: 01 September 2022 Date Date:
 Document No. Page: 4 of 2
Revision Date
Department: Pathology 01 September
2022
Section: Effective Date:
Revision No. 01September 2022

Policy Title:
STANDARD OPERATING PROCEDURE IN RHEUMATOID FACTOR (RF)
**Other substances not tested may interfere. Clinical diagnosis should not be
made on findings of a single test result, but should integrate both clinical and laboratory
data.

XI. REFERENCES
 Singer J.M., Plotz, C.M. Amer. J. Med. 21:888-895 (1956)
 Singer J.M., Plotz, C.M. Amer. J. Med. Assoc.168 : 180 (1959)
 Dorner R.W. Et. Al. ClinicachimicaActa. 1987:167:1-21
 Wolfe F. et al. Arthritis and Rheumarism 1991;34:951 - 960

Prepared By: Reviewed By: Noted by: Approved by:

MARIE ANGELICA HABALO, RMT KHRISTINE T. MORENO, RMT CARINA P. VILLAMAYOR,MD,FPSP Roy O. Arceo, MD, MHA, FPSO-HNS
Date: 01 September 2022 Date: 01 September 2022 Date Date: