| Project/Area Number |
16590337
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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 |
Parasitology (including Sanitary zoology)
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
KATAKURA Ken Hokkaido University, Graduate School of Veterinary Medicine, Professor, 大学院・獣医学研究科, 教授 (10130155)
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| Co-Investigator(Kenkyū-buntansha) |
TSUBOI Takafumi Ehime University, Cell-Free Science and Technology Research Center, Professor, 無細胞生命科学工学研究センター, 教授 (00188616)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2004: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | antigens / cutaneous leishmaniasis / diagnosis / Leishmania (leishmania) major / Leishmania (Viannia) panamensis / serum / Time of Flight Mass Spectrometry (TOF-MS) / Western blot / 血清診断 / Leishmania(L.)major / Leishmania(V.)panamensis / Leishmania(L.)tropica / レコンビナント蛋白 / Leishmania amazonensis / Leishmania mexicana / Leishmania panamensis |
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| Research Abstract |
The present study was aimed to identify immunodiagnostic antigens specific to each of cutaneous, muco-cutaneous and visceral leishmaniasis and develop a rapid serological diagnostic kit for cutaneous leishmaniasis using the wheat germ cell-free system as a novel protein biosynthetic method. In murine leishmaniasis, immunoblot patterns were different with Leishmania species inoculated such as Leishmania (Leishmania) donovoni, L.(L.) major and L.(L.) amazonensis although antibodies common to all Leishmania species used were produced. In terms of cutaneous leishmaniasis in the New World, antigens of approximately 120,95,85 and 75 kDa were frequently recognized by sera from patients with cutaneou leishmaniasis in endemic areas of Leishmania (Viannia) panamensis in Ecuador. These antigens were analysed by Matrix Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry (TOF-MS). The resultant candidate proteins detected were elongation factor 2(94 kDa), heat shock protein 83(80 kDa) and heat shock protein 70(72 kDa) of L.(V.) braziliensis, a close related species to L.(V.) panamensis. Production of these recombinant proteins using the wheat germ cell-free system is underway. In terms of cutaneous leishmaniasis in the Old World, we found that the main causative agent of outbreak of cutaneous leishmaniasis in southern Pakistan was L.(L.) major followed by L.(L.) tropica. Western blot analysis of sera from these patients is underway.
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