Mycoplasma hyopneumoniaeの種特異抗原に関する研究 Studies on specific antigens of Mycoplasma hyopneumoniae.



    • 森, 康行 モリ, ヤスユキ



Mycoplasma hyopneumoniaeの種特異抗原に関する研究


Studies on specific antigens of Mycoplasma hyopneumoniae.


森, 康行


モリ, ヤスユキ











Mycoplasma hyopneumoniae is one of the most common pathogens of swine respiratory disease.・ Mycoplasma pneumonia of swine(NPS) caused by this organism is a widespread, chronic and economically important disease throughout the world. Although intensive works on the pathogenesis, diagnosis and prevention of this disease have been continued since the discovery of M.hyopneumoniae by Mare and Switzer, many problems are still remaining to be resolved.Especially,little is known about surface antigenic structure and it’s functions. For example, it is well known that growth of mycoplasmas is inhibited by antibody, however,antigen(s) perticipates in this growth inhibition is not still identified in any mycoplasma species. On the other hand, many attempts have been made to develop a useful serodiagnostic test for MPS in different laboratories, employing complement fixation(CF), indirect hemagglutination and enzyme-linked immunosorbent assay (ELISA). etc.. However, until recently none of these techniques was completely satisfactory because of cross reaction between M.hyopneumoniae and other swine mycoplasmas. I It is thought that antigen analysis and use of specific antigen(s) of M.hyopneumoniae might be very effective to develop a more specific and sensitive diagnostic tests. Furthermore, analysis and identification of antigen(s) participating in antibody mediated growth inhibition may give a promising candidate for component vaccine against MPS. For these purposes, I analyzes the serological cross-reactivity among swine mycoplasmas and antibody response of swine against M.hyopneumoniae were produced for analysis of antigens and applied them to the serological diagnosis and identification of M.hyopneumoniae. The results obtained are as follows:1. Mycoplasmas solubilized with Tween 20 were used as antigen. Tween 20-treated antigens of M.hyopneumoniae were subjected to column chromatography on Sephacryl S-300 for purification. Of five main peaks, the second gave the most reliable results with reduction of non-specific reactions. 2. In cross-reacting ELISA using the 2nd peak of Tween 20 treated antigens, less prominent but detectable cross-reactions were observed between M.hyopneumoniae antigen and anti-M.flocculare serum. And it was demonstrated that the predominant crossreactive antigens were components of M.hyopneumoniae with molecular weight of 74 kilodalton(Kd) and 53Kd. 3. It was indicated that 96Kd, 70Kd, 46Kd and 38Kd antigenic components of M.hyopneumoniae and 67Kd, 56Kd and 23Kd antigens of N.flocculare were specific for respective species. 4.Antibodies against 96Kd, 76Kd, 70Kd, 53Kd, 46Kd and 38Kd antigens of M.hyopneumoniae were detected in serum from pigs experimentally infected with M.hyopneumoniae. Of these anti-bodies those against 46Kd antigen reacted early andwere most consistently detected during experimental infection. 5. Twelve clones of hybridoma producing monoclonal antibody against M.hyopneumoniae were established and nine of these monoclonal antibodies were specific for M.hyopneumoniae. 6. Three monoclonal antibodies (E-7-1-4,J-168-1 and J-49-7) specifically agglutiniated M.hyopneumoniae and recognized the glycoprotein-like components on the surface of M.hyopneumoniae, Of these monoclonal antibodies, E-7-1-4 inhibited the growth ofM.hyopneumoniae. 7. Monoclonal antibody (14-4-4) against 46Kd antigen of M.hyopneumoniae could be applied successfully to the ELISA double sandwich method to detect swine antibodies against 46Kd antigen of M.hyopneumoniae. This procedure gave high specificity and sensitivity. 8.In the experimental infection with M.hyopneumoniae, anti-bodies were detected by the double sandwich ELISA antibodies persisted foe longer period than those detected by the CF test. 9.In the field application of this ELISA using nonclonal antibody, of the 1461 serum samples collected from breeding swine herd in Japan, 1285(88%) had antibodies against M.hyopneumoniae. 10.Novel identification technique for M.hyopneumoniae was developed by employing agglutinating monoclonal antibody (E-7-1-4).


Hokkaido University(北海道大学). 博士(獣医学)


  1. 緒言 / p1 (0005.jp2)
  2. 第1章 Mycoplasma hyopneumoniae抗原の分析 / p5 (0009.jp2)
  3. 要旨 / p5 (0009.jp2)
  4. 材料と方法 / p5 (0009.jp2)
  5. 成績 / p8 (0012.jp2)
  6. 考察 / p13 (0017.jp2)
  7. 第2章 M.hyopneumoniaeに対するモノクローナル抗体の作出 / p15 (0019.jp2)
  8. 要旨 / p15 (0019.jp2)
  9. 材料と方法 / p15 (0019.jp2)
  10. 成績 / p20 (0024.jp2)
  11. 考察 / p30 (0034.jp2)
  12. 第3章 抗M.hyopneumoniaeモノクローナル抗体を用いた豚マイコプラズマ肺炎の血清学的診断と分離菌株の迅速同定 / p34 (0038.jp2)
  13. 要旨 / p34 (0038.jp2)
  14. 材料と方法 / p34 (0038.jp2)
  15. 成績 / p35 (0039.jp2)
  16. 考察 / p44 (0048.jp2)
  17. 総括 / p47 (0051.jp2)
  18. 参考文献 / p49 (0053.jp2)
  19. 謝辞 / p55 (0059.jp2)
  20. 英文抄録 / p56 (0060.jp2)


    • 8000000081418
  • DOI(NDL)
  • 本文言語コード
    • jpn
  • NDL書誌ID
    • 000000245524
  • データ提供元
    • 機関リポジトリ
    • NDLデジタルコレクション