A sandwich method of enzyme-immunoassay. II. Quantification of rheumatoid factor.

A sandwich enzyme-immunoassay (EIA) has been applied to the determination of the rheumatoid factor (RF). This non-competeitive assay comprises 3 steps: 1) the RF to be assayed is extracted for the biological medium by an immunosorbent of aggregated IgG linked to cellulose; 2) the solid phase is then incubated with the enzyme-labeled aggregated IgG; 3) the enzymatic activity of the immunosorbent is then measured with a suitable chromogenic reagent. This activity is a direct function of the amount of RF to be assayed. This assay gave reproducible results in the range 0.5-50.0 IU/ml. A good agreement was obtained between the EIA and the Waaler-Rose test but no correlation was obtained with the latex slide-test. This assay permits a quantitation of RF with a good reproducibility (coefficient of variation in the range of 10% for moderately elevated values) and thus allows a closer follow-up of patients. The results do not depend on the interpretation of the technician performing the test, which can be easily automated. Finally, it may detect some RF devoid of agglutinating activity.

[Clinical applications of enzyme-immunoassay of rheumatoid factor]. / Applications cliniques du dosage enzymo-immunologique du facteur rhumatoïde.

The authors describe a method of enzymo-immunological estimation of rheumatoid factor. This estimation, of non-competitive type, includes 3 stages : the rheumatoid factor is first extracted from the serum by fixation on the aggregated IgG linked to cellulose powder; then its presence is revealed by fixation of an oxidase IgG glucose conjugate on the solid phase; finally, the enzyme activity linked to the solid phase is measured. This activity is directly related to the concentration of rheumatoid factor present. The estimation, carried out on 2 050 patients including 114 with rheumatoid arthritis, is well correlated with technic of the Rose-Waaler reaction. Furthermore, the greater sensitivity, the better reproducibility and the quantitative character of this method might improve the supervision of patients with rheumatoid arthritis.

[Enzymo-immunologic determination of the class of antiglobulin factors]. / Détermination enzymo-immunologique de la classe des facteurs antiblobuliniques.

The authors describe an enzymo-immunologic method of determination of the class of rheumatoid factors (RF). This technic includes 3 main stages : 1) extraction of RF by fixation on aggregated rabbit IgG previously adsorbed on the walls of a plastic tube; 2) recognition of RF linked to the solid phase by anti-IgG addition, and addition of human IgA and IgM linked to glucose oxidase; 3) revelation of the enzyme activity linked to the solid phase : this enzyme activity directly depends on the levels of RF to be estimated. The results are expressed as a percentage of the fixation of a reference serum obtained from a patient with rheumatoid arthritis (RA). This serum presents an antiglobulin activity in the 3 major classes of immunoglobulins. The coefficients of variation of intra- and intersystem reproductibility lay between 5,6 and 17.8%. The specificity of the estimation was controlled by the localisation of the antiglobulin activity of the sera after chromatography on Sephadex G200 and by estimation of RF after their absorption by an aggregated IgG immunoadsorbant. This technic was applied to the identification of RF in RA (90 cases); in diseases not related to RA and in control subjects : the sensitivity of this method appears greater than that of technics previously described (immuno-fluorescence, Rose-Waaler reaction) in fact, only 12,2% of sera of patients with RA remain negative with this new test. However, this greater sensitivity does not seem to have altered the clinical specificity of the test : in fact, only 8,4% of the sera of patients with diseases unrelated to RA are positive by this method, which, moreover, may be explained by the age of these subjects (average age 71.5 years).

Non-excretory multiple myeloma.

The clinical and laboratory features in five patients with non-excretory myeloma are described including plasma cell immunofluorescence and, in two cases, ultrastructure. Findings are compared with those in similar patients previously reported, and those in excretory disease. Clinical, haematological and biochemical features were similar to those found in excretory myeloma, showing differences only in relation to the absence of serum or urinary monoclonal immunoglobin. Cellular cytological and ultrastructural features allowed no differentiation from excretory cells. Within this non-excretory group distinction between secretory and non-secretory myeloma cells is possible on the basis of immunofluorescence. Differing patterns of intermittent excretion occurred in three of these patients.