Rose-Waaler test using a rapidly prepared serum fraction.

The use of a serologically active fraction of serum in parallel with whole serum, in testing for the sheep cell agglutinating factor in rheumatoid arthritis, was suggested by Svartz and Schlossmann (1953) and by Ziff, Brown, Badin, and McEwen (1954). Claims of higher sensitivity and specificity have been re-emphasized in more recent publications (Ziff, Brown, Lospalluto, Badin, and McEwen, 1956; Svartz and Schlossmann, 1957). Since 1956, sheep cell agglutination tests have been performed in this laboratory by Ziff's method, using absorbed serum, euglobulin fraction, and inhibition titres as criteria (Rose and Whillans, 1957). This experience confirms the view of Hess (1956) that, while the technique is laborious, it is not outside the scope of a laboratory doing routine serological tests. Although the exact nature of the rheumatoid agglutinating factor is not yet elucidated, it is usually considered to migrate in an electrical field mainly with the gamma globulins (Lamont-Havers, 1955; Svartz and Schlossmann, 1955; Lospalluto and Ziff, 1956). Thulin (1955) considered it to be distributed over the faster moving gamma and beta globulins, while Heller, Kolodny, Lepow, Jacobson, Rivera, and Marks (1955) classified it mainly as a beta-lipoprotein. Recently Franklin, Kunkel, Muller-Eberhard, and Holman (1957) presented evidence from ultracentrifugal studies for a fast moving gamma globulin being responsible for the serological activity in rheumatoid arthritis and a similar view has been put forward by Svartz (1957). The preparation of the euglobulin fraction used in Ziff's method is a time consuming process, so that a search was made for a simpler method of preparing the active fraction. In view of the above it appeared to us that a gamma globulin fraction would be desirable. Serum fractions consisting mainly of gamma globulins can be prepared, using ammonium sulphate for salting out, by prolonged standing. However, the rapid and simple method of Wolfsohn, Cohn, Calvary, and Ichiba (1948) gives a complete precipitate in less than one hour, and when haemagglutination titres using this fraction and Ziff's euglobulin showed good parallelism it was decided to compare the two fractions further.