Complement fixation test using the antigen prepared from the nervous tissue of adult mice infected with poliomyelitis virus Lansing type.

The technique of complement fixation test as a measure for the serological diagnosis of neurotropic virus infections was first put in practice by Casals and Palacios (1) in the year 1941, and confirmed of its practical diagnostic value by numerous follow-ups made after it. In spite of repeated attempts, the application of this technique for the diagnosis of poliomyelitis had not been successful. The main reason for this failure was attributed to the fact that the preparation of antigen in pure and highly concentrated form was difficult. In the year 1947, H. S. Loring, S. Raffel and J. C. Anderson (2) inoculated cotton rats with rodent pathogenic poliomyelitis virus Lansing strain. The central nervous tissue of inoculated animals was put into an emulsion which was subsequently centrifuged by ultra-centrifugation and the supernatant fluid of nitrogen content 10-5g/ml was obtained. Using this supernatant as an antigen, they performed complement fixation tests with the sera of cotton rats and monkeys immunized with the virus of Lansing strain and with the convalescent sera of poliomyelitis patients. The reactions were positive and they proved the possibility of applying this reaction for the diagnosis of poliomyelitis which had hitherto been considered impracticable. Later in the year 1951, J. Casals, P. K. Olitsky and R. O. Anslow (3, 4) reported that they obtained positive results in the complement fixation tests made with the antigen prepared by following Casals' acetone-ether method (5) except that the strain used was Lansing type MEF which had been increased of its virulence by the passages through new born mice. Their antigen showed the complement fixation reaction not only with the immune serum of rodent pathogenic strain but also with the immune serum of monkey pathogenic Brunhilde strain up to a certain degree. Following the above, O. Lahelle (6) proved the specific occurrence of the complement fixation reaction employing the antigen prepared by acetone-ether treatment from the nervous tissue of cotton rats of 1 to 3 days after birth which had been inoculated intracerebrally with the virus of Lansing type. Further, M. Pollard (7, 8) carried out the complement fixation test using the antigen prepared by methanol purification from the nervous tissues of cotton rats which had been inoculated with the virus of Lansing type, but the results obtained were not satisfactory. Thus, he employed the purification method of ultra-centrifugation which yielded