Instruction Manual B02135

Brizuela-Lab.

rabbit gamma globulin teeth. Grieble and collaborators (7) and

Rose Ragan Latex Janeff (10) in 1966 and 1970, respectively, present statistics
For the detection of the Rheumatoid factor by the reaction of regards. The last of those mentioned, using the new method in
Rose Ragan, in diluted serum. object holder with rabbit gamma globulin latex, comes to the conclusion-
session that their results are between high sensitivity and low
INTRODUCTION specificity, such as the latex test and high specificity and the
low sensitivity of the Waaer-Rose.
The serum of patients suffering from Rheumatoid Arthritis contains
the so-called rheumatoid factors (RF). Macroglobulins that
ELEMENTS OF THE SYSTEM
they could be considered as antibodies, taking into account their
chemical composition and its relation to gamma globulins of the type Provided
19 S. 1. Rabbit gamma globulin latex antigen: Suspension of
Rheumatoid factors have the property of reacting agglutinatingly latex particles of 0.25 microns in diameter coated with
Nando or precipitating with human gamma globulin and that of some... rabbit gamma globulin, brought to an aggregated state.
our animals (2), (8), (15). 2. Glycine buffer: Aminoacetic acid 240 mmol/L, hydroxide of
The presence of these factors in arthritics varies, according to the researchers. 90 mmol/L
players from 70 to 90% (11), (17), (19). Although they can be Positive Control: Stabilized control corresponding to positive.
found also in other processes, such as syphilis (14), tuberculosis dad 2 crosses (++)
(21), lepra (5), enfermedades hepáticas (1), endocarditis bacteriana 4. Negative Control: Solution control free of agglutinins.
(23) and chronic bronchitis (3), the frequency is undoubtedly significant.
again smaller. REQUIRED MATERIAL
It is important to note that, according to some researchers, the FR. It is Not provided
present in about 3.5% of healthy people (17), having it Glass plate.
found in blood donors in percentages ranging from 4 to 7.6 2. Clock or Timer.
Its frequency is higher in healthy people over 65. 3. Rod.
years (8), (12). 4. Light Source.
On the other hand, it is absent in a significant percentage of nurses. 5. Suitable volumetric material.
arthritis, although in these cases it can be found in the fluid 6. Glass tubes.
synovial.
The most common reactions for the detection of the rheumatic factor- STABILITY AND CONSERVATION
latex fixation test, which employs The provided reagents are stable until the expiration date.
action of Rose Ragan, who uses gamma indicated on the packaging stored between (2º-10º C). Do not freeze.
rabbit globulin.
Milgrom and collaborators (13) demonstrated the existence of three SAMPLE
distinct factors that together constitute what is called F.R.: Serum will be used, which must be inactivated for 10 minutes at
one of them reacts only to human gamma globulin, the 60º C. This procedure removes the globulin ter-
second it faces human gamma globulin and that of molable 11S (8), which combines with gamma globulin in state
rabbit, while the third reacts only to the of aggregation, producing inhibition of the reaction. With the inacti-
rabbit gamma globulin. the phenomenon of pro-zone is avoided, which causes false results
This would explain the behavior of the serum from arthritic patients. negative data in arthritic sera, when working with low doses
things that can react positively with the latex test or dilutions of the same, (2); (6); (20). If the determination is not
vice versa, or result positively in both cases. The sample can be kept for a week if done on the day.
In other words, the use of rabbit gamma globulin and the preserved between 2-10°C.
human in simultaneous reactions, increases the percentage of
positivity in the detection of rheumatoid arthritis. PROCEDURE
Qualitative Technique
USE FOR WHICH IT IS INTENDED Bring the reagents to room temperature. Shake the reagent.
For the detection of the Rheumatoid factor by the Rose reaction Latex-Globulin before use. Make a 1/8 dilution of the sample.
Ragan. inactivated train (see Sample) placing, in a tube, 50 µl of sample-
try 0.35 ml of glycine buffer.
FOUNDATION On a glass plate place:
When in 1922 Meyer describes the influence of serum on patients Sample 1/8 1 drop (50 µl)
of rheumatoid arthritis on the agglutination of erythrocytes of Latex-Globulin Reagent 1 drop (50 µl)
ram by the amphitrochanter; laid the foundations of serology of the Mix with a stick or glass rod until obtaining a
A.R.; but it remained without practical application until 1940 when uniform suspension, extend it to a diameter of 20
Waaler studied this problem thoroughly. It is in 1948 that Rose, Ragan gently balance the plate for 2 minutes.
and collaborators (17) perfected the reaction (hence it is called Observe the presence or absence of agglutination.
also Waaler-Rose reaction) and obtained encouraging results microscopic. Reading is facilitated by placing a source
All (80% of positives sick with A.R. and 3..,5 of false positives of light under the plate.
in healthy controls).
In 1952, Winblad established that the serum of patients with R.A. pre-
Quantitative Technique
he had an affinity for gamma globulin. He proposed a test based on
erythrocytes treated with tannin and absorbed with gamma globulin
Positive sera can be titrated. For this, in 5 tubes of
Human. In 1956, Singer and Plotz replaced the red blood cells
by latex particles, this constitutes the so-called fixed test glass
latex coagulation, a procedure that is carried out in tubes and results in Place 0.2 ml of glycine buffer in each tube.
the one with the highest sensitivity that exists for the detection of the F.R. (not Take 0.2 ml of the 1/8 dilution and transfer it to tube No. 1. Mix.
must be confused with the latex plate test). It is followed by the modification- and transfer 0.2 ml to tube No. 2 and so on until the last one
Rhelms' proposal in 1957 (16) suggests working with droplets, with a tube. The obtained dilutions will be: 1/16–1/32–1/64–1/128–
considerable savings in time and reagent. Singer, in 1958 (19), 1/256.
adapt the test implementation to plates. Test each dilution as indicated in the qualitative technique.
The simplification that occurred in the tests that use gamma
human globulin, was reflected in the parallel for the corresponding

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 Instruction Manual B02135
Brizuela-Lab.

EXPRESSION OF RESULTS Helmer R. Et al (1971): "Globulins resembling rheumatoid factor"

If no agglutination occurred with the inactivated serum, it will be reported: in serum of the aged.” Am. J. of Med. 35. 175.
Rose Ragan Latex: Non-reactive serum. Huskisson et al (1971): 'Synovial fluid Waaler Rose and lates'
Otherwise, the greater dilution that produces agglutination us test. Ann. Rhem. Dis. 30:67.
indicate the positivity title. For example: Reactive serum 1/32. Janeff J. (1970): “A new slide test for rheumatoid arthritis: com-
Rose Ragan Latex: Reactive serum dilution... comparison with Waaler-Rose and latex slide tests. Arth and Rheum.
Reactive results are considered as negative reactions. 13:193.
up to the dilution 1/8, agglutination at dilution 1/16 is considered Klein F. et al. (1966): 'Characterization of two different'
doubtful and frankly positive reaction when agglutination is obtained agglutinators in the latex fixation test occurring in normal human
from the dilution 1/32 onwards. sera. Immunology 10:87
Larsen R. (1972): Family studies in systemic lupus
QUALITY CONTROL erythematosus" J. Chron. Dis. 25: 191.
Since statistical quality control in serology is not easy- Milgrom F. Et al (1965): “Multiplicity of rheumatoid factor” arth,
practical mind, what is proposed is a control of reactivity and Reum. 8.203.
from the latex suspension, for which a positive serum is supplied Peltier et al (1959): 'The presence of rheumatoid factor in sera
diluted as a control, which can be analyzed simultaneously with from patients with syphilis. [Link] Rhem. 2.1.
each set of reactions that are practiced. Pike R. et al (1963): "Species specificity of globulins in reactions"
with rheumatoid arthritis serum." J. Immunol. 9:740.
LIMITATIONS AND CARE Rheins M. Et al (1957): “A modification of the latex fixation test
Pollution with detergents: The presence of detergents for the study of rheumatoid arthritis." J. Lab. And Clin. Med. 50:
it can alter the agglutination process, so it will be necessary to 113.
take the precaution of ensuring that the plate is free of Rose H.; Ragan C. et al (1948): Differential agglutination of normal
these products. and sensitized sheep erythrocytes by sera of patients with rheuma-
toid arthritis.” Proc. Soc. Exp. Biol: and Med. 68:1.
rabbit stays out of the refrigerator for extended periods Singer J. and Plotz C. (1956): "Latex fixation test: Application to
goads, a marked decrease in reactivity occurs. serologic diagnosis of rheumatoid arthritis. Am. J. Med. 21:888.
The freezing of the latex suspension causes alterations Singer J. (1958): 'Slide latex fixation test: a simple screening'
irreversible in it, rendering it permanently unusable. method for the diagnosis of rheumatoid arthritis." J.A.M.A. 168:
180
BIBLIOGRAPHY Singer J. (1961): 'The latex fixation test in rheumatic diseases'.
Atwater E. Et al (1963): “The lates-fixation in patient with liver Am.J. of Med. 31:766.
disease, Ann. Int. Med. 58: 419. Singer J. et al (1962): “The presence of antigamma globulin factors
Bernard G. et al (1962): "The reaction of rheumatoid factor and in sera of patients with active pulmonary tuberculosis.” Ann Int.
complement with gamma globulin coated latex. Immunology 88: Med. 56:545.
750. Waller M. et al (1961): "Evaluation of rheumatoid factor"
Bonomo L. Et al (1966): “Anti gamma globulin and antinuclear test. Arthr. and Rheum. 4:579.
factors in chronic bronchitis”. Am J. of clin. Path. 45 : 313. Williams et al (1962): "Rheumatoid factor, complement and
Caplan H. (1971): “Latex-fixation test to screen blood donors”. N. conglutinin aberrations in patients with sub acute bacterial endo-
Enl. JH. Med 285.295 carditis”.J. Clin. Invest. 41:666.
Cathcart C. Et al (1961): "The relationship of the latex fixation test"
to the clinical and serologic manifestation of leprosy. [Link]. PRESENTATION
31:758 Package for 40 determinations. COD B02135
Chuy-Tsai Cheg et al (1971): “Interference by Ciq in latex slide test
for rheumatoid factor.” Ann. Int. Med. 75: 683.
Grieble H. et al (1969): "The serologic diagnosis of rheumatoid" Product made by Laboratorios W. Brizuela S.A.
arthritis. The latex–rabitgamma globulin fixation test” J. Chron. Av. Figueroa Alcorta 123/139 5000–Córdoba (Argentina)
Dis.21:667. info@[Link]
Product authorized by ANMAT No. 6505
Technical Director: Biochemist Marcelo Brizuela

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