レプトスピラ症の血清診断への蛍光抗体法間接法の応用に関する研究

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書誌事項

タイトル別名
  • Studies of Application of Indirect Fluorescent Antibody Technique for Serodiagnosis of Leptospirosis
  • レプトスピラ症の血清診断への蛍光抗体法間接法の応用に関する研究-1-免疫ウサギ血清を用いての基礎実験
  • レプトスピラショウ ノ ケッセイ シンダン エ ノ ケイコウ コウタイホウ カ
  • 1. Basal Experiments with Immune Rabbit Sera
  • 1. 免疫ウサギ血清を用いての基礎実験

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An indirect fluorescent anti-leptospiral antibody technique (FLAi) was studied to detect the antibody in immune rabbit sera. Seven serotypes of leptospira found in Japan were used as the antigens. The fluorescein-labelled antibody used was “Dried Fluorescein-conjugated Goat Antiserum to Rabbit 7S Globulin Laboratory Reagent (Hyland)”. For fixation of the antigen, Kirkpatrik's solution, mixture of ethylalcohol, ether and formalin (6: 3: 1) was found to be fittest. Anti-leptospiral immune sera were prepared by injecting rabbits with 0.1% formalin killed leptospiral suspensions which were made of fresh cultures in Korthof's medium, stored at-20C until use.<BR>The procedure of FLAi was as follows: Leptospiral antigen was smeared on slide glass, fixed, dried, and layered with each of 2-fold serial dilutions of a test serum, sensitized at 37C for an hour in a wet room, and washed several times first with PBS and then with purified water. After drying by the fan, the smear was layered with the fluorescein-labelled antibody, sensitized at 37C for an hour in a wet room following by washing and drying as mentioned above. It was embedded in PBS-glycerin, examined under the fluorescence microscope. The result was evaluated by BV excitation with No.3 filter (Y51).Agglutination was carried out by the Schiifner-Mochtar's microscopic agglutination (MA) method.<BR>Both FLAi and MA gave the highest titers against the homologous serotype antigen respectively, though FLAi showed higher cross reactivity as compared with the case of MA. A rabbit serum immunized with Reiter strain of Treponema produced positive reaction with 3 of 6 antigens at 40-fold serum dilution, while a normal rabbit serum was perfectly negative. In the rabbit infected with a vilulent copenhageni, the antibody was shown by FLAi and MA using the homologous antigen at least for 8 weeks, though FLAi titer tended to lower than MA titer. The reactivity to heterologous antigen was demonstrated by FLAi but not by MA. The mixture of various serotypes may be suitable for epidemiological survey of leptospirosis as the antigen of FLAi.<BR>The detection of leptospira by FLAi technique from urine of pyrogenes-carrying mouse and from blood of copenhageni infected guinea pig was attempted, good stainability of the organisms was demonstrated.<BR>In laboratory possessing the fluorescent antibody technique, it may be possible to detect anti-leptospiral antibody by FLAi supplying no reagents but the leptospira antigen.

収録刊行物

  • 感染症学雑誌

    感染症学雑誌 54 (8), 365-373, 1980

    一般社団法人 日本感染症学会

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