On the macroscopical rickettsia agglutination test on the hollow slide-glass.

Abstract

Since it was revealed that the causative agent for. the murine and epidemic typhus, scrub typhus, Rocky Mountain Spotted fever, trech fever and other diseases of similar character were all of rickettsial diseases, an attempt has been made to establish the agglutination test with each homologous rickettsia as an antigen not only for the specific diagnosis (1, 2) and differentiation (3, 4) but also for the study on the antigenic structure (3, 5) of each rickettsia.In earlier days, the antigen for rickettsia agglutination test (R.A.T.) was prepared from the following materials, emulsion of eggs (1) from the infected ticks with Ricket-tsia (R.) rickettsii, each emulsion of the intestines (2, 3, 5, 7, 8) of the infected lice with each R. prowazekii, R. typhi and R. quintana, peritoneal washing (7) of the white rats in-fected with typhus fever, suspension of slide cell culture (10) with R. prowazekii, and emul-sion Of infected lung tissue (9, 11, 19) of mice or rats with typhus fever. And the judgement was made under microscope (1, 2, 4, 14, 16) on the preparates stained by Giemsa's solution. However it was noted that these methods did not afford a satisfactory result.In 1932, Zinsser (3) has designed a special capillary test-tubes charged with 0.025 cc. each of the antigen and the test serum placed in a water bath at 40°C. for 2 to 5 hours and left standing in an ice-chamber overnight for inspection by naked eyes on the agglutination. Since then such process (4, 5, 7, 8) and modified method (11) have been in practice by workers in despite of much difficulties in preparation of the antigen before Cox (6) has, in 1938, published of his yolk sac culture method to result in an abundantt harvest much easily of pure rickettsia-body.The remarkable progress has thus been made in R.A.T, when Plotz (12) reported of the method of reading macroscopically the agglutination of rickettsia antigen pre-pared by Cox-Craigie's method, namely, 0.25 cc, of the antigen mixed with the same amount of the serum to be tested in test-tubes in the way similar to the Widals test with Salmonella typhi, incubated at 42°C. for 4 hours and left standing in an ice-chamber (4°C) for 16 to 18 hours and then read by naked eyes. This method requires too much amount of the antigen to be in practice generally by the physician, therefore, a simplified orientating method on slide-glasses with a loopful amount of the antigen was recommended by Fitz-Patrick (21), Castaneda (9) and Kitaoka (20) .Furthermore the attempt was made by us to quantitative estimations of R. A. T. with each two-fold dilution of the test serum by mixing with the equal amount of thee antigen as minimal as a drop on a hollow slide-glass and read the result by naked eyes to differentiate the typhus fever group from the other acute infectious diseases, as well as the epidemic typhus from the murine.