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Pasteurella multocida 에 대한 간이적혈구 응집반응과 적혈구의 안정화에 관한 연구

Studies on the Simplified Hemagglutination Reaction to Pasteurella multocida and the Stabilization of Erythrocytes

대한수의학회지 1970년 10권 1호 p.1011 ~ 1023
정영건 (  ) - 영남대학교 축산대학

이학철 ( Lee Hak-Cheul ) - 영남대학교 축산대학
김교준 (  ) - 충남대학교 농과대학

Abstract


Recently Carter(1952) reported the capsule antigens of Pasteurella nultocida could be divided into four serological types A,B,C and D by means of precipitation tests. Subsequently he showed that the most sensitive for identification of these types involved the use of capsule substance adsorbed by erythrocytes in hemagglutination test. It may be somewhat difficult to conduct the hemagglutination test in small laboratory, because relatively large amounts of antisera and erythrocytes of the human O type are required for the test.
A simple method for serological typing of P. multocida was the slide agglutination test employed by Little et al. (1943) and Namioka et al. (1962), but this method is still in controversy.
The author tried adapting Carter’s hemagglutination method to the slide method so called “micromethod technique”, and studied on the stabilization of erythrocytes for use of slide hemagglutination to P. multocida although many invesigators reported the stabilization of erythrocytes.
The results obtained are summarized as follows:
1. A simplified method (slide method) for capsule typing of the organism was developed by adapting Carter’s hemagglutination reaction(tube method). Antibody-containing serum can be diluted serially on Boerner’s microtest slide with capillary or serological pipetts with a considerable accuracy. The slide reaction can be carried out with case on the slide by adding 0.05㎖ of antigensensitized erythrocytes suspension diluted to one percent on 0.05㎖ of serially diluted antibodycontaining sera, and the final result can be read after 60 minutes at the room temperature (15℃).
2. It is difficult to determine superiority of inferiority between the slide method and the tube method on the pattern of the reaction of hemagglutination.
3. The pH range of 6.6 to 8.3 is optimal for the slide hemagglutination reaction.
4. The antigen-sensitization against erythrocytes at 37℃ is optimal for the slide hemagglutination.
5. Both the doses and concentration of antigen do not influence the antigen-adsorbing capacity of erythrocytes.
6. The reduction of antigen-sensitizing hours does not influence the antigen-adsorbing capacity of erythrocytes even 30 minutes.
7. The tannic acid treatment against formalinized and non-formalinized erythrocytes showed no effect on the reaction of hemagglutination.
8. The erythrocytes preserved at 4℃ in the ACD solution do not decrease the reactivity on the reaction of hemagglutination for 60 days, while they begin slight hemolysis 30 days after preserving.
9. The stable preparation of erythrocytes can be obtained by treating the cells at 37℃ for 20 hours with from 4 to 8 percent of formalin in saline or buffer. These cells can be preserved at 4℃ for more than 8 months experimented without hemolysis. With low concentration of formalin, the cells were not sufficiently stabilized resulting in the hemolysis after short period of preservation at 4℃.
10. The erythrocytes treated with 16 percent of formalin remain constantly or increase the reactivity for the reaction of hemagglutination. On the contrary, the cells treated with I to 8 percent of formalin decrease the reactivity.
11. There is no difference between nontreated fresh erythrocytes and the erythrocytes preserved in the ACD solution on the reactivity against the hemagglutination, and the erythrocytes treated with 16 percent of formalin showed the reactivity of higher level than that of the above two kinds of erythrocytes.
12. There is no difference between the saline and the isotonic buffier solution on the reaction of hemagg1utination.

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