1990 Fiscal Year Annual Research Report

病原因子を指標にした下痢原因菌の迅速な免疫学的検出法の開発

Research Project

Project/Area Number	01870020
Research Institution	Research Institute fo Microbial Diseases, Osaka University
Principal Investigator	三輪谷俊夫大阪大学, 微生物病研究所, 教授 (60029759)
Co-Investigator(Kenkyū-buntansha)	坂本雅子財団法人阪大微生物病研究会, 係長有田美知子大阪大学, 微生物病研究所, 助手 (10127178) 余明順大阪大学, 微生物病研究所, 助手 (70093482) 山本耕一郎大阪大学, 微生物病研究所, 助手 (30158274) 本田武司大阪大学, 微生物病研究所, 助教授 (60029808)
Keywords	腸炎ビブリオ / 毒素原性大腸菌 / 抗体 / ELISA
Research Abstract	I.検出法の開発 (1)抗体の整備: これまでにわれわれが単離精製できた種々な病原因子(定着因子,蛋白毒素)に対するポリクロ-ナルおよびモノクロ-ナル抗体を完備することができた。 (2)検出系の開発:ELISAをはじめ、Elek変法(沈降法)の応用や凝集法(プロテインAブドウ球菌,ラテックスなど)を,種々の病原因子の検出系として応用した。これらの中で最も実用性の高いものは、ELISA法およびElek変法と思われた。また、ごく最近入手できたエバフィルム(化学的にシ-ト上に抗体を結合させる)を利用した系も試み中で,有望な見透しを得つつある。 II.検出系の応用 ELISAを用いると、予備的な動物実験系ではあるが病原因子を直接検体(下痢便)中に証明できることがわかり、下痢症の迅速診断に応用できる可能性が示された。現在,大阪空港検疫所を中心にヒト下痢便について、ELISA等の有用性を検討中である。これまでに得られたLT産生性毒素性大腸菌および腸炎ビブリオ(TDH)を対象とした検討結果によると、かなりの精度でそれぞれの存在の有無を判定できたが、一方で非特異的な陽性反応も見られたので、さらに検体(ヒト下痢便)の調整法の工夫を行っている。 III.マニュアルの作成分離菌の産生する病原因子の検出に有用な免疫学的な検出系を多数確立することができたので、これらの実施法についてのマニュアルを作成した。留学生等も利用できるように和文のみならず英文で解説したマニュアルも用意した。

Research Products

(18 results)

All Other

All Publications (18 results)

[Publications] Honda,T.,et al.: "Production and characterization of monoclonal antibodies to a pilus colonization factor (colonization factor antigen III) of human enterotoxigenic <Escherichia>___ー <coli>___ー." Infect.Immun.57. 3452-3457 (1989)
[Publications] Yoh,M.,et al.: "Homogeneity of a hemolysin (VhーrTDH) produced by clinical produced by clinical and environmental isolates of <Vibrio>___ー <hollisae.>___ー" FEMS Microbiol.Lett.61. 171-176 (1989)
[Publications] Tsuji,T.,et al.: "Charge heterogeneity of heatーlabile enterotoxins from human enterotoxigenic <Escherichia>___ー <coli>___ー." FEMS Microbiol.Lett.61. 1-6 (1989)
[Publications] Yoh,M.,et al.: "Comparative amino acid sequence analysis of hemolysins produced by <Vibrio>___ー <hollisae>___ー and <Vibrio>___ー <parahaemolyticus>___ー." J.Bacteriol.171. 6859-6861 (1989)
[Publications] Honda,T.,et al.: "Production of monoclonal antibody against a hemolysin (VpーTRH) produced by <Vibrio>___ー <parahaemolyticus>___ー." FEMS Microbiol.Lett.68. 167-170 (1990)
[Publications] Tsuji,T.,et al.: "A heatーlabile enterotoxin (LT) purified from chicken enterotoxigenic <Escherichia>___ー <coli>___ー is identical to poruine LT." FEMS Microbiol.Lett.67. 329-332 (1990)
[Publications] Honda,T.,et al.: "Properties of a hemolysin related to the thermostable direct hemolysin produced by a Kanagawa phenomenon negative,clinical isolate of <Vibrio>___ー <parahaemolyticus>___ー." Can.J.Microbiol.36. 395-399 (1990)
[Publications] Tsuji,T.,et al.: "Comparison of coligenoid formation by B subunits of procine and human <Escherichia>___ー <coli>___ー heatーlabile" FEMS Microbiol.Lett.69. 299-304 (1990)
[Publications] Joya,J.E.,et al.: "Demonstration of enterotoxigenic <Escherichia>___ー <coli>___ー in diarrheic broiler chickes." Eur.J.Epidemiol.6. 88-90 (1990)
[Publications] Yamamoto,K.,et al.: "Twoーstep processing for activation of cytolysin/hemolysin of <Vibrio>___ー <cholerae>___ー 01,El Tor;nucleotide sequence of the structural gene and (hlyA) and characterization of the processed products." Infect.Immun.
[Publications] Tsuji,T.,et al.: "Detection and purification of the free A subunit of heatーlabile enterotoxin produced by enterotoxigenic <Escherichia>___ー <coli>___ー." FEMS Microbiol.,Lett.
[Publications] Honda,T.,et al.: "Characterization of a new thermostable direct hemolysin produced by a Kanagawa phenomenonーnegative clinical isolate of <Vibrio>___ー <parahaemolyticus>___ー." J.Gen.Microbiol.
[Publications] Honda,T.,et al.: "Production of monoclonal antibody against a hemagglutinin/protease of <Vibrio>___ー <cholerae>___ー nonー01." FEMS Microbiol.Lett.
[Publications] 三輪谷俊夫: "ビフィズス2(250頁のうち9頁)" (財)日本ビフィズス菌センタ-, 9 (1989)
[Publications] 三輪谷俊夫: "日本内科学会雑誌 79(3)(700頁のうち5頁)" 日本内科学会, 5 (1990)
[Publications] 三輪谷俊夫: "感染症学雑誌 64(7)(1262頁のうち7頁)" 感染症学会, 7 (1990)
[Publications] 三輪谷俊夫: "腸炎ビブリオ第III集(監修三輪谷俊夫,大橋誠)(362頁のうち8頁)" 近代出版, 8 (1990)
[Publications] 三輪谷俊夫: "消化器外科13(6)(1560頁のうち1頁)" へるす出版, 1 (1990)

1990 Fiscal Year Annual Research Report

病原因子を指標にした下痢原因菌の迅速な免疫学的検出法の開発

Principal Investigator

三輪谷俊夫大阪大学, 微生物病研究所, 教授 (60029759)

Research Products

[Publications] Honda,T.,et al.: "Production and characterization of monoclonal antibodies to a pilus colonization factor (colonization factor antigen III) of human enterotoxigenic <Escherichia>_ー <coli>_ー." Infect.Immun.57. 3452-3457 (1989)

[Publications] Yoh,M.,et al.: "Homogeneity of a hemolysin (VhーrTDH) produced by clinical produced by clinical and environmental isolates of <Vibrio>_ー <hollisae.>_ー" FEMS Microbiol.Lett.61. 171-176 (1989)

[Publications] Tsuji,T.,et al.: "Charge heterogeneity of heatーlabile enterotoxins from human enterotoxigenic <Escherichia>_ー <coli>_ー." FEMS Microbiol.Lett.61. 1-6 (1989)

[Publications] Yoh,M.,et al.: "Comparative amino acid sequence analysis of hemolysins produced by <Vibrio>_ー <hollisae>_ー and <Vibrio>_ー <parahaemolyticus>_ー." J.Bacteriol.171. 6859-6861 (1989)

[Publications] Honda,T.,et al.: "Production of monoclonal antibody against a hemolysin (VpーTRH) produced by <Vibrio>_ー <parahaemolyticus>_ー." FEMS Microbiol.Lett.68. 167-170 (1990)

[Publications] Tsuji,T.,et al.: "A heatーlabile enterotoxin (LT) purified from chicken enterotoxigenic <Escherichia>_ー <coli>_ー is identical to poruine LT." FEMS Microbiol.Lett.67. 329-332 (1990)

[Publications] Honda,T.,et al.: "Properties of a hemolysin related to the thermostable direct hemolysin produced by a Kanagawa phenomenon negative,clinical isolate of <Vibrio>_ー <parahaemolyticus>_ー." Can.J.Microbiol.36. 395-399 (1990)

[Publications] Tsuji,T.,et al.: "Comparison of coligenoid formation by B subunits of procine and human <Escherichia>_ー <coli>_ー heatーlabile" FEMS Microbiol.Lett.69. 299-304 (1990)

[Publications] Joya,J.E.,et al.: "Demonstration of enterotoxigenic <Escherichia>_ー <coli>_ー in diarrheic broiler chickes." Eur.J.Epidemiol.6. 88-90 (1990)

[Publications] Yamamoto,K.,et al.: "Twoーstep processing for activation of cytolysin/hemolysin of <Vibrio>_ー <cholerae>_ー 01,El Tor;nucleotide sequence of the structural gene and (hlyA) and characterization of the processed products." Infect.Immun.

[Publications] Tsuji,T.,et al.: "Detection and purification of the free A subunit of heatーlabile enterotoxin produced by enterotoxigenic <Escherichia>_ー <coli>_ー." FEMS Microbiol.,Lett.

[Publications] Honda,T.,et al.: "Characterization of a new thermostable direct hemolysin produced by a Kanagawa phenomenonーnegative clinical isolate of <Vibrio>_ー <parahaemolyticus>_ー." J.Gen.Microbiol.

[Publications] Honda,T.,et al.: "Production of monoclonal antibody against a hemagglutinin/protease of <Vibrio>_ー <cholerae>_ー nonー01." FEMS Microbiol.Lett.

[Publications] 三輪谷俊夫: "ビフィズス2(250頁のうち9頁)" (財)日本ビフィズス菌センタ-, 9 (1989)

[Publications] 三輪谷俊夫: "日本内科学会雑誌 79(3)(700頁のうち5頁)" 日本内科学会, 5 (1990)

[Publications] 三輪谷俊夫: "感染症学雑誌 64(7)(1262頁のうち7頁)" 感染症学会, 7 (1990)

[Publications] 三輪谷俊夫: "腸炎ビブリオ第III集(監修三輪谷俊夫,大橋誠)(362頁のうち8頁)" 近代出版, 8 (1990)

[Publications] 三輪谷俊夫: "消化器外科13(6)(1560頁のうち1頁)" へるす出版, 1 (1990)

1990 Fiscal Year Annual Research Report

病原因子を指標にした下痢原因菌の迅速な免疫学的検出法の開発

Principal Investigator

三輪谷 俊夫 大阪大学, 微生物病研究所, 教授 (60029759)

Research Products

[Publications] Honda,T.,et al.: "Production and characterization of monoclonal antibodies to a pilus colonization factor (colonization factor antigen III) of human enterotoxigenic <Escherichia>___ー <coli>___ー." Infect.Immun.57. 3452-3457 (1989)

[Publications] Yoh,M.,et al.: "Homogeneity of a hemolysin (VhーrTDH) produced by clinical produced by clinical and environmental isolates of <Vibrio>___ー <hollisae.>___ー" FEMS Microbiol.Lett.61. 171-176 (1989)

[Publications] Tsuji,T.,et al.: "Charge heterogeneity of heatーlabile enterotoxins from human enterotoxigenic <Escherichia>___ー <coli>___ー." FEMS Microbiol.Lett.61. 1-6 (1989)

[Publications] Yoh,M.,et al.: "Comparative amino acid sequence analysis of hemolysins produced by <Vibrio>___ー <hollisae>___ー and <Vibrio>___ー <parahaemolyticus>___ー." J.Bacteriol.171. 6859-6861 (1989)

[Publications] Honda,T.,et al.: "Production of monoclonal antibody against a hemolysin (VpーTRH) produced by <Vibrio>___ー <parahaemolyticus>___ー." FEMS Microbiol.Lett.68. 167-170 (1990)

[Publications] Tsuji,T.,et al.: "A heatーlabile enterotoxin (LT) purified from chicken enterotoxigenic <Escherichia>___ー <coli>___ー is identical to poruine LT." FEMS Microbiol.Lett.67. 329-332 (1990)

[Publications] Honda,T.,et al.: "Properties of a hemolysin related to the thermostable direct hemolysin produced by a Kanagawa phenomenon negative,clinical isolate of <Vibrio>___ー <parahaemolyticus>___ー." Can.J.Microbiol.36. 395-399 (1990)

[Publications] Tsuji,T.,et al.: "Comparison of coligenoid formation by B subunits of procine and human <Escherichia>___ー <coli>___ー heatーlabile" FEMS Microbiol.Lett.69. 299-304 (1990)

[Publications] Joya,J.E.,et al.: "Demonstration of enterotoxigenic <Escherichia>___ー <coli>___ー in diarrheic broiler chickes." Eur.J.Epidemiol.6. 88-90 (1990)

[Publications] Yamamoto,K.,et al.: "Twoーstep processing for activation of cytolysin/hemolysin of <Vibrio>___ー <cholerae>___ー 01,El Tor;nucleotide sequence of the structural gene and (hlyA) and characterization of the processed products." Infect.Immun.

[Publications] Tsuji,T.,et al.: "Detection and purification of the free A subunit of heatーlabile enterotoxin produced by enterotoxigenic <Escherichia>___ー <coli>___ー." FEMS Microbiol.,Lett.

[Publications] Honda,T.,et al.: "Characterization of a new thermostable direct hemolysin produced by a Kanagawa phenomenonーnegative clinical isolate of <Vibrio>___ー <parahaemolyticus>___ー." J.Gen.Microbiol.

[Publications] Honda,T.,et al.: "Production of monoclonal antibody against a hemagglutinin/protease of <Vibrio>___ー <cholerae>___ー nonー01." FEMS Microbiol.Lett.

[Publications] 三輪谷 俊夫: "ビフィズス2(250頁のうち9頁)" (財)日本ビフィズス菌センタ-, 9 (1989)

[Publications] 三輪谷 俊夫: "日本内科学会雑誌 79(3)(700頁のうち5頁)" 日本内科学会, 5 (1990)

[Publications] 三輪谷 俊夫: "感染症学雑誌 64(7)(1262頁のうち7頁)" 感染症学会, 7 (1990)

[Publications] 三輪谷 俊夫: "腸炎ビブリオ第III集(監修 三輪谷 俊夫,大橋 誠)(362頁のうち8頁)" 近代出版, 8 (1990)

[Publications] 三輪谷 俊夫: "消化器外科13(6)(1560頁のうち1頁)" へるす出版, 1 (1990)

三輪谷俊夫大阪大学, 微生物病研究所, 教授 (60029759)

[Publications] Honda,T.,et al.: "Production and characterization of monoclonal antibodies to a pilus colonization factor (colonization factor antigen III) of human enterotoxigenic <Escherichia>_ー <coli>_ー." Infect.Immun.57. 3452-3457 (1989)

[Publications] Yoh,M.,et al.: "Homogeneity of a hemolysin (VhーrTDH) produced by clinical produced by clinical and environmental isolates of <Vibrio>_ー <hollisae.>_ー" FEMS Microbiol.Lett.61. 171-176 (1989)

[Publications] Tsuji,T.,et al.: "Charge heterogeneity of heatーlabile enterotoxins from human enterotoxigenic <Escherichia>_ー <coli>_ー." FEMS Microbiol.Lett.61. 1-6 (1989)

[Publications] Yoh,M.,et al.: "Comparative amino acid sequence analysis of hemolysins produced by <Vibrio>_ー <hollisae>_ー and <Vibrio>_ー <parahaemolyticus>_ー." J.Bacteriol.171. 6859-6861 (1989)

[Publications] Honda,T.,et al.: "Production of monoclonal antibody against a hemolysin (VpーTRH) produced by <Vibrio>_ー <parahaemolyticus>_ー." FEMS Microbiol.Lett.68. 167-170 (1990)

[Publications] Tsuji,T.,et al.: "A heatーlabile enterotoxin (LT) purified from chicken enterotoxigenic <Escherichia>_ー <coli>_ー is identical to poruine LT." FEMS Microbiol.Lett.67. 329-332 (1990)

[Publications] Honda,T.,et al.: "Properties of a hemolysin related to the thermostable direct hemolysin produced by a Kanagawa phenomenon negative,clinical isolate of <Vibrio>_ー <parahaemolyticus>_ー." Can.J.Microbiol.36. 395-399 (1990)

[Publications] Tsuji,T.,et al.: "Comparison of coligenoid formation by B subunits of procine and human <Escherichia>_ー <coli>_ー heatーlabile" FEMS Microbiol.Lett.69. 299-304 (1990)

[Publications] Joya,J.E.,et al.: "Demonstration of enterotoxigenic <Escherichia>_ー <coli>_ー in diarrheic broiler chickes." Eur.J.Epidemiol.6. 88-90 (1990)

[Publications] Yamamoto,K.,et al.: "Twoーstep processing for activation of cytolysin/hemolysin of <Vibrio>_ー <cholerae>_ー 01,El Tor;nucleotide sequence of the structural gene and (hlyA) and characterization of the processed products." Infect.Immun.

[Publications] Tsuji,T.,et al.: "Detection and purification of the free A subunit of heatーlabile enterotoxin produced by enterotoxigenic <Escherichia>_ー <coli>_ー." FEMS Microbiol.,Lett.

[Publications] Honda,T.,et al.: "Characterization of a new thermostable direct hemolysin produced by a Kanagawa phenomenonーnegative clinical isolate of <Vibrio>_ー <parahaemolyticus>_ー." J.Gen.Microbiol.

[Publications] Honda,T.,et al.: "Production of monoclonal antibody against a hemagglutinin/protease of <Vibrio>_ー <cholerae>_ー nonー01." FEMS Microbiol.Lett.

[Publications] 三輪谷俊夫: "ビフィズス2(250頁のうち9頁)" (財)日本ビフィズス菌センタ-, 9 (1989)

[Publications] 三輪谷俊夫: "日本内科学会雑誌 79(3)(700頁のうち5頁)" 日本内科学会, 5 (1990)

[Publications] 三輪谷俊夫: "感染症学雑誌 64(7)(1262頁のうち7頁)" 感染症学会, 7 (1990)

[Publications] 三輪谷俊夫: "腸炎ビブリオ第III集(監修三輪谷俊夫,大橋誠)(362頁のうち8頁)" 近代出版, 8 (1990)

[Publications] 三輪谷俊夫: "消化器外科13(6)(1560頁のうち1頁)" へるす出版, 1 (1990)