| Project/Area Number |
13671841
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Ophthalmology
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| Research Institution | The University of Tokushima |
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Principal Investigator |
SHIOTA Hiroshi Graduate School, Institute of Health Biosciences, Professor, 大学院ヘルスバイオサイエンス研究部, 教授 (20035736)
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| Co-Investigator(Kenkyū-buntansha) |
EGUCHI Hiroshi Tokushima University Hospital, Department of Ophthalmology, Assistant Professor, 医学部・歯学部附属病院, 講師 (30314868)
TAJIKA Tomoyuki Tokushima Municipal Hospital, Department of Ophthalmology, Head, 眼科, 医長 (30325253)
SHINOMIYA Kayo The University of Tokushima, Graduate School Institute of Health Biosciences, Instructor, 大学院ヘルスバイオサイエンス研究部, 助手 (80322262)
HATA So Hata Eye Clinic, Director, 院長 (50343306)
INOUE Sadako Asada General Hospital, Department of Ophthalmology, Head, 眼科, 医長
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2002: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2001: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | keratomycosis / gene diagnosis / genome / PCR / PCR-SSCP / Candida / Fusarium |
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| Research Abstract |
Keratomycosis must be diagnosed and treated early. However, it is hard to make the diagnosis and the isolation of fungus from the corneal lesion takes 5 to 10 days. Moreover, the identification of the fungus and drug sensitivity tests take several weeks. We developed a rapid diagnostic method to identify the fungus within one night using recent molecular biological technique. Also, we established a guide for the treatment of keratomycosis after analyzing our 62 cases.
1) The development of a rapid diagnostic method for keratomycosis: Based on data from the Gene Bank and many literatures we selected primers to target 18S rRNA gene for medically common fungi, cutinase gene for Fusarium, 18S rRNA gene of different portion of the previous gene for Aspergillus and cytochrome P450L1A1 demethylase gene for Candida. Using patients' samples, standard fungi and identified fungi from patients, we tested our selected primers and found that all these primers worked very good. When we carried out PCR
… More
-SSCP using patients' samples and our selected primers, we found that we could identify fungi whether they were Fusarium, Aspergillus or Candida within one night. These results show that the identification of gungi is possible within one night if our rapid diagnostic method using PCR-SSCP is taken. In the past it took several weeks. This early identnification of gungi is also very helpful in selecting the anti-fungal drug from the medical point of view.
2) Animal experiment: Candida albicans was inoculated into the rabbit corneas. Eleven days after the inoculation the corneal lesion was scraped. The PCR and ordinally isolation methods were compared using the scraped materials which shows better results. Our above PCR method showed much better results than the ordinally isolation method in the animals, too.
3) Establishment of a guide for the treatment of keratomycosis: Our 62 cases of keratomycosis were analyzed carefully and the following guide lines were established. Pimaricin is the drug of choice for the treatment of keratomycosis. When clinical conditions are not improved after 5 to 7 days with pimaricin, other anti-fungal drugs must be switched or combined with pimaricin. Less
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