エキノコックス症免疫診断法(国際標準診断、早期診断、治癒判定法)の実用化

2002 Fiscal Year Annual Research Report

• Project/Area Number: 12557024
• Research Institution: Asahikawa Medical College
• Principal Investigator: 伊藤 亮 旭川医科大学, 医学部, 教授 (70054020)
• Co-Investigator(Kenkyū-buntansha): 山崎 浩 旭川医科大学, 医学部, 助教授 (00138207) ; 中谷 和宏 旭川医科大学, 医学部, 助教授 (70109388) ; 高後 裕 旭川医科大学, 医学部, 教授 (10133183) ; 中尾 稔 旭川医科大学, 医学部, 助手 (70155670) ; 藤本 佳範 旭川医科大学, 医学部, 講師 (90292127)
• Keywords: エキノコックス症 / 多包虫症 / 単包虫症 / 有鉤嚢虫症 / 遺伝子組み換え診断抗原 / 血清学的鑑別診断 / ミトコンドリアDNA / 遺伝子多型

Research Abstract

本研究における主要研究テーマは、現在地球規模で流行が深刻化しているエキノコックス症(多包虫症、単包虫症)に関する信頼性の高い免疫診断法(国際標準診断、早期診断、治癒判定法)の開発と実用化である。
(1)現在、国際的に最も信頼性が高いと評価されている多包虫症特異診断抗原Em18を遺伝子組み換え抗原(RecEm18)として作成し、その過程でEm18の生化学的特性の解析がなされた。世界各国で別個に検索されてきた診断抗原の中でEm18のB cell epitope活性が最も高いことが判明した(Sako et al. 2002. J Clin Microbiol 40,2760-2765)。(2)フランスとのブラインドテストによる共同研究から、RecEm18を用いる血清診断法はフランスにおける多包虫症と単包虫症を100%鑑別できることが判明した(Ito et al. 2002. J Clin Microbiol 40,4161-4165)。現在、フランス、スイスにおいて認定された300例以上の多包虫症例の血清学的確認要請を受けている。(3)単包虫症の血清診断法についても遺伝子組み換えAntigen Bを作製した(Mamuti et al. 2002. Clin Diag Lab Immunol 9,573-576;Mamuti et al. in prep.)。(4)血清診断上、最も交差反応が高い致死的寄生虫疾患、有鉤嚢虫症に関する遺伝子組み換え抗原を作製した(Sako et al. 2000. J Clin Microbiol 38,4439-4444)。血清診断上鑑別を要する多包虫症、単包虫症、有鉤嚢虫症すべてに関する遺伝子組み換え抗原作製に成功し、国際的な評価を得た。現在、合成ペプチド抗原作製を試みている。(5)北海道を中心に、主治医から直接相談を受けた多包虫症疑診例において、Em18抗原を用いる血清検査で多包虫症と他の疾患を術前に100%鑑別し、術前確定血清診断法を確立した。国内での現行の血清検査が確定検査として機能していないことから、Em18を用いる血清検査の導入が望まれる。(6)ミトコンドリアDNA解析により、遺伝子診断法の開発が可能になった。遺伝子解析、遺伝子診断研究も含め、これらの研究成果は現在印刷中を含め国際誌に原著論文総説29編、国際会議録14編、著書5篇として発表した。和文総説、報告書も22編発表した。

Research Products

• [Publications] Ito, A.: "Perspective on control options for Echinococcus multilocularis"Parasitology. 127(in press). (2003)
• [Publications] Ito, A.: "Review : Echinococcosis and cysticercosis in Asia : evaluation of the modern technology for epidemiological study"Southeast Asian Journal of Tropical Medicine and Public Health. 34(in press). (2003)
• [Publications] Yamasaki, H.: "Cysticercosis/taeniosis : recent advances in serological and molecular diagnoses"Southeast Asian Journal of Tropical Medicine and Public Health. 34(in press). (2003)
• [Publications] Ito, A.: "Multiple genotypes of Taenia solium-ramifications for control"Acta Tropica. (in press). (2003)
• [Publications] Nakao, M.: "The abbreviated stop codon U found in the mitochondrial genome of the tapeworm Taenia solium"Journal of Parasitology. (in press). (2003)
• [Publications] Ito, A.: "Control of echinococcosis and cysticercosis : a public health challenge to international cooperation in China"Acta Tropica. (in press). (2003)
• [Publications] Ito, A.: "Detection and control of zoonotic cestode infections : immunodiagnostic and molecular approaches"Trends in Parasitology. (in press). (2003)
• [Publications] Ito, A.: "Development of Em18-immunoblot and Em18-ELISA for special diagnosis of alveolar echinococcosis"Acta Tropica. 85. 173-182 (2003)
• [Publications] Ito, A.: "Recent advances in basic and applied science for the control of taeniasis/cysticercosis"Southeast Asian Journal of Tropical Medicine and Public Health. 33(in press). (2002)
• [Publications] Ito, A.: "Collaboration for the control of cysticercosis/echinococcosis in Vietnam"Southeast Asian Journal of Tropical Medicine and Public Health. 33(in press). (2002)
• [Publications] Margono, S.S.: "Taenia solium taeniasis/cysticercosis in Papua, Indonesia in 2001 : detection of human worm carriers"Journal of Helminthology. 77. 39-42 (2003)
• [Publications] Sato, M.O.: "Evaluation of tongue inspection and serology for diagnosis of Taenia solium cysticercosis in swine : usefulness of ELISA using purified glycoproteins and recombinant antigen"Veterinary Parasitology. 111. 309-322 (2003)
• [Publications] Ito, A.: "Dogs as alternative intermediate hosts of Taenia solium in Papua(Irian Jaya), Indonesia confirmed by highly specific ELISA and immunoblot using native and recombinant antigens and mitochondrial DNA analysis"Journal of Helminthology. 76. 311-314 (2002)
• [Publications] Ito, A.: "Evaluation of an enzyme-linked immunosorbent assay(ELISA) with affinity-purified Em18 and ELSIA with recombinant Em18 for differential diagnosis of alveolar echinococcosis : results of a blind test"Journal of Clinical Microbiology. 40. 4161-4165 (2002)
• [Publications] Ito, A.: "Human Taenia in NOD-scid mice : how can we use such an animal model?"Proceedings of the 10^<th> International Congress of Parasitology. 179-183 (2002)
• [Publications] Ito, A: "Taeniasis/cysticercosis in Papua(Irian Jaya), Indonesia"Proceedings of the 10^<th> International Congress of Parasitology. 207-211 (2002)
• [Publications] Yamasaki, H.: "DNA differential diagnosis of human taeniid cestodes using restriction fragment length polymorphism and base excision sequence scanning T-base system"Proceedings of the 10^<th> International Congress of Parasitology. 189-203 (2002)
• [Publications] Sato, M.O.: "Comparison of traditional tongue examination and serology for diagnosis of swine cysticercosis : usefulness of ELISA using native and recombinant antigens"Proceedings of the 10^<th> International Congress of Parasitology. 195-199 (2002)
• [Publications] Nakao, M.: "The complete mitochondrial DNA sequence of the cestode Echinococcus multilocularis (Cyclophyllidea : taeniidae)"Mitochondrion. 1. 497-509 (2002)
• [Publications] Yamasaki, H.: "DNA differential diagnosis of human taeniid cestodes by base excision sequence scanning thymine-base reader analysis with mitochondrial genes"Journal of Clinical Microbiology. 40. 3818-3821 (2002)
• [Publications] Ito, A.: "Serologic and molecular diagnosis of zoonotic larval cesode infections"Parasitology International. 51. 221-235 (2002)
• [Publications] Margono, S.S.: "Multiple cysticercus nodules in skin and brain in a Balinese woman : a case report"Medical Journal of Indonesia. 11. 169-173 (2002)
• [Publications] Sako, Y.: "Alveolar echinococcosis : characterization of diagnostic antien Em18 and serological evaluation of recombinant Em 18"Journal of Clinical Microbiology. 40. 2760-2765 (2002)
• [Publications] Yamasaki, H.: "Characterization and expression of the Fasciola gigantica cathepsin L gene"International Journal for Parasitology. 32. 1031-1042 (2002)
• [Publications] Nakao, M.: "A phylogenetic hypothesis for the distribution of two genotypes of the pig tapeworm Taenia solium worldwide"Parasitology. 124. 657-662 (2002)
• [Publications] Manuti, W.: "Usefulness of hydatid cyst fluid of Echinococcus granulosus developed in mice with secondary infection for serodiagnosis of cystic echinococcosis in humans"Clinical and Diagnostic Laboratory Immunology. 9. 573-576 (2002)
• [Publications] Ito, A.: "Cry wolf!"Trends in Parasitology. 18. 47-48 (2002)
• [Publications] Ito, A.: "Comparison of the antigenicity of protoscoleces and microvesicles of Echinococcus multilocularis prepared from rats"Journal of Helminthology. 75. 355-358 (2001)
• [Publications] Subahar, R.: "Taenia solium infection in Irian Jaya(West Papua), Indonesia : a pilot serological survey of human and porcine cysticercosis in Jayawijaya District"Transactions of the Royal Society of Tropical Medicine and Hygiene. 95. 388-390 (2001)
• [Publications] Jiang, L.: "Immunodiagnostic differentiation of alveolar and cystic echinococcosis using ELISA test with 18-kDa antigen extracted from Echinococcus protoscoleces"Transactions of the Royal Society of Tropical Medicine and Hygiene. 95. 285-288 (2001)
• [Publications] Wang, Q.: "Investigation of risk factors for development of human hydatidosis among households raising livestock in Tibetan areas of western Sichuan province"Chinese Journal of Parasitology and Parasitic Diseases. 19. 93-96 (2001)
• [Publications] Margono, S.S.: "The problem of taeniasis and cysticercosis in Irian Jaya, Indonesia"Medical Journal of Indonesia. 10. 110-114 (2001)
• [Publications] Margono, S.S.: "Cysticercosis in Indonesia : epidemiological aspects"Southeast Asian Journal of Tropical Medicine and Public Health. 32. 79-84 (2001)
• [Publications] Ito, A.: "NOD-scid mouse as an experimental animal model for cysticercosis"Southeast Asian Journal of Tropical Medicine and Public Health. 32. 85-89 (2001)
• [Publications] Okamoto, M.: "Molecular variation of Taenia solium in the world"Southeast Asian Journal of Tropical Medicine and Public Health. 32. 90-93 (2001)
• [Publications] Sako, Y.: "Recent advances in serodiagnosis for cysticercosis"Southeast Asian Journal of Tropical Medicine and Public Health. 32. 98-104 (2001)
• [Publications] Ito, A.: "Differential serodiagnosis of cystic and alveolar echinococcosis using native and recombinant antigens in Japan"Southeast Asian Journal of Tropical Medicine and Public Health. 32. 111-115 (2001)
• [Publications] Ito, A.: "Neurocysticercosis and echinococcosis in Asia : Recent advances in the establishment of highly reliable differential serodiagnosis for international collaboration"Southeast Asian Journal of Tropical Medicine and Public Health. 31. 16-20 (2000)
• [Publications] Nakao, M.: "Mitochondrial genetic code in cestodes"Molecular and Biochemical Parasitology. 111. 415-424 (2000)
• [Publications] Sako, Y.: "Molecular characterization and diagnostic value of Taenia solium low-molecular-weight antigen genes"Journal of Clinical Microbiology. 38. 4439-4444 (2000)
• [Publications] Craig, P.S.: "An epidemiological and ecological study of human alveolar echinococcosis transmission in south Gansu, China"Acta Tropica. 77. 167-177 (2000)
• [Publications] Wandra, T.: "Resurgence of cases of epileptic seizures and burns associated with cysticercosis in Assologaima, Jayawijaya, Irian Jaya, Indonesia, 1991-95"Transactions of the Royal Society of Tropical Medicine and Hygiene. 94. 46-50 (2000)
• [Publications] 伊藤 亮: "特集 人畜共通感染症 肺エキノコックス症"呼吸. 22. 56-60 (2003)
• [Publications] 伊藤 亮: "特集テーマ:今日常診療で注目すべき原虫症、寄生虫症 エキノコックス症"JIM. 13(in press). (2002)
• [Publications] 畠山優一: "肝単包虫症の1切除例"手術. 56. 819-823 (2002)
• [Publications] 伊藤 亮: "技術講座 単包虫症、多包虫症の免疫学的診断"Medical Technology. 30. 97-103 (2002)
• [Publications] 伊藤 亮: "寄生虫学講座におけるエキノコックス症、有鉤嚢虫症、ライム病についての研究総括"旭川医科大学研究フォーラム. 2(2). 14-21 (2001)
• [Publications] 伊藤 亮: "新興・再興寄生虫症(有鉤嚢虫腫御、エキノコックス症)の世界における現状 10,59-66"臨床環境医学. 10. 59-66 (2001)
• [Publications] 伊藤 亮: "エキノコックス症の疫学と血清診断"Current Concepts in Infectious Disease. 20. 18-19 (2001)
• [Publications] 伊藤 亮: "執筆者からの返事"北海道医学雑誌. 76. 367-368 (2001)
• [Publications] 伊藤 亮: "旭川医科大学におけるエキノコックス症研究の現状と日本におけるエキノコックス症の問題点"北海道医学雑誌. 76. 3-8 (2001)
• [Publications] 伊藤 亮: "新興・再興寄生虫病としてのエキノコックス症、有鉤嚢虫症"岐阜県医師会医学雑誌. 14. 35-46 (2001)
• [Publications] 伊藤 亮: "エキノコックス症を取り巻く諸問題"旭川医科大学研究フォーラム. 2(1). 13-19 (2001)
• [Publications] 伊藤 亮: "エキノコックス症(多包虫症)血清診断の問題点"Infection Control. 10. 44-46 (2001)
• [Publications] 伊藤 亮: "エキノコックス症-世界・日本での現状、診断・治療・予防-"北海道プライマリケア研究会会報. 19. 31-41 (2001)
• [Publications] Ito, A.: "In : Progress of Medical Parasitology in Japan. Chapter III. Session 6. Immunology in cestode infections (2) Immunity to the larval cestodes"Meguro Parasitology Museum, Tokyo(in press). (2003)
• [Publications] Ito, A.: "Taenia solium Cysticercosis. Chapter 5. Mitochondrial DNA of Taenia solium : from basic to applied science"CABI Press, Oxson, UK. 47-55 (2002)
• [Publications] Singh, G.: "Taenia solium Cysticercosis. Chapter 12. Taenia solium taeniasis and cysticercosis in Asia"CABI Press, Oxson, UK. 111-127 (2002)
• [Publications] Ito, A.: "Cestode Zoonoses : Echinococcosis and Cysticercosis-An Emergent and Global problem. Neurocysticercosis in Asia : serology/seroepidemiology in humans and pigs"IOS Press, Amsterdam. 25-31 (2002)
• [Publications] Ito, A.: "Cestode Zoonoses : Echinococcosis and Cysticercosis-An Emergent and Global problem. Differential serodiagnosis for alveolar echinococcosis by Em18-immunoblot and Em18-ELISA in Japan and China"IOS Press, Amsterdam. 147-155 (2002)
• [Publications] 皆川知紀: "北海道感染症対策マニュアル"北海道保健福祉部保健予防課. 242 (2001)
• [Publications] 伊藤 亮: "シンプル微生物学(改訂第3版)"南江堂. 343-354 (2000)