| Budget Amount *help |
¥8,000,000 (Direct Cost: ¥8,000,000)
Fiscal Year 1985: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1984: ¥6,000,000 (Direct Cost: ¥6,000,000)
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| Research Abstract |
The immunodiagnosis for Pneumocystis carinii (Pc) pneumonia are very difficult for detection of their antibodies under the immunosuppresive conditions, therefore, the methods for identification of antigenic substances, so called circulating antigen of Pc in the infected sera are trying. However, these methods and criteria for diagnosis are not yet determined. Comparative studies on experimental and patients sera have been done by the several methods. The results are as follows.
1. Enzyme-labeling IgG antibody was preparated by Periodate method and Fab' antibody was used by Malaimide method. And experimental rat sera were examined by Sandwich technique of ELISA. Under the condition of 128 <micro> g protein/ml for plate coating and X4 dilution of examined sera showed the best results by Periodate method, and 16 <micro> g protein/ml, X10 dilution sera by Malaimide method.
2. In the cases of patients sera, about 30 % among normal sera showed positive under the same conditions. These ELISA technique seems to be very sensitive but the latent infection cases showed positive, too.
3. The crude antigen was purified by Sephacryl S 200 and Polyacrylamide gel, and hyper immune sera were prepared with this purified antigen. About 20 % casesamong experimental Pc rat sera showed negative results by Ouchterlony method.
4. 552 patients sera of 382 cases were examined by Ouchterlony method. The sera within one week after bleeding showed parallel results with clinical findings, but all freezed stock sera in the period of one month over showed negative results. It was recognized that antigenic activity of patients sera was decreased even in freezing condition. Radioimmunoassay and the approach of monoclonal antibody are not yet developed. In the moment, Ouchterlony was most suitable method for diagnosis.
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