結核補體結合性抗原 관한 硏究 = The Study of Complement Fixing Antigen for the Diagnosis of Tuberculosis

다국어 초록 (Multilingual Abstract)

The author has observed the specificity and antigenecity of two kinds of antigen, one of which is carbohydrate moiety and the other is the protein.
Measurement of hemolysin titre: The immunized serum of rabbits by sheep RBC was used as haemolysin, and it was used in three fold concentration of the complete hemolysis titrated preliminary test. Hemolysin was always used after deciding its titre at preliminary tet. The sheep RBC suspension was used by 3 per cent.
The dosage of antigen:1/2 amount of the maximum haemolysin does, determined by the result of preliminary test of the author's two kinds of antigen, was used.
Complement: Complement was obtained by the heart puncture of healthy guinea pigs. The two fold amount of the complement titre at preliminary titration was used in this experiment.
Test sera were all supplied from the from the patients who were recieving treat-ment in the T. B. centre attached to the Ministry of Health and Welfare of Korea. These sera were inactivated by heating at 56℃ for 30 min., and when they were needed preservation, they were stored in refrigerator at 40℃± and were reheated at 56℃ for 15min. just before submitting to the experiment.
Procedures of making antigens: C antigen-Very virulent human tubercle bacillus strain C is cultivated for 40 days on Sauton's non-protein media, and washed several times with saline on the filter papers. After dessication, the organisms are suspended in saline at the rate of 10 mg/ml and submitted to the rapid manipulation of freezing and thawing between -20℃ and 20℃;after 20 times of this manipulation, it is centrifuged at 8,000 rpm for 30 min, and its supernatant is picked up as C antigen.
P-antigen: The same dried strain as used in making the C antigen is heated by steam at 100℃ for 30min., filtered through Seitz filter, into this filtered fluid added 40% trichlor acetic acid at the rate of 4 per cent, and set this fluid at 4℃±all day long and overnight. After then, its sedimented substance is isolated, suspended in 1% trichlor acetic acid solution, centrifuged by 3,000 rpm for 10 min., and its sedimented substance is re-picked up. This procedures are repeated twice more quite the same way, and into the finally got substance .by centrifugation is added 0.3% KH₂PO₄in approximate amount, and, after enough mixing, it is centrifuged so as to wash away the trichlor acetic acid.
This manipulation of washing must be repeated three timesm and then the sediment is dried without heating, its powder is dissolved in phosphate buffered solution (pH 7.0) at the rate of 1.0 mg/ml, then it is the stock solution of P antigen, (the dried powder was got at the rate of 10.1mg from 2,500 ml of filtered fluid, and the total nitrogen amount was 13.34% by microki jeldahl method).
The complement-fixation-reaction was carried out in T. B. serum using P-and C-antigens made by the author, and following results were obtained:
1) Both P-and C-antigens had neither anti-complement, nor exclusive hemolytic action.
2) Antigenecity of C-antigen is weak, and becomes weaker by heating and by preserving long time, while that of P-antigen being considerable high and never influenced by heating or by long period of preservation that is, not decreased even by heating at 60℃ for 30minutes, nor by preservation for 3 days at 37℃, nor for one month at room temperature.
3) Proving of antibody for the complement-fixation-reaction in lung T.B. serums is difficult immediately after drawing of blood, but it gradually increases as time goes, about 7 days later it becoming the maximum, and ,then after 12 days, again, it disappears.
4) Antibody detecting rate of lung T.B. is paralell with the degree of disease severity, but in utmost severe cases it's detection is rather difficult.
5) The rate of non-specific positive reaction by P-antigen with syphilis serum is low.
6) But with leprosy serum the non-specific positive rate of P-antigen is considerably high, especially in nodular case it is twice that of nervous leprosy.
7) The non-specific positive rate of P-antigen to the serum of healthy person is very low.

목차 (Table of Contents)

• I. 緖 論
• Ⅱ. 實驗方法
• 1. 豫備實驗
• 2. 抗原의 作成
• 3. 本試驗
• Ⅲ. 實驗成績
• 1. 抗原에 대한 成績
• 2. 結核補體結合性抗體의 出現移動
• 3. 結核補體結合反應成績
• 4. 癩患者에 對한 成績
• 5. 梅毒患者에 對한 成績
• 6. 健康人에 對한 成績
• Ⅳ. 總 括
• Ⅴ. 結 論