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ID 11264
Eprint ID
11264
FullText URL
Title Alternative
Highly Sensitive Detection of Cucumber Mosaic Virus by Using Fluorogenic Substrate or Enzyme-Amplified Assay in Enzyme-Linked Immunosorbent Assay
Author
Maeda, Takanori
Sako, Nobumichi
Inouye, Narinobu
Abstract
Monoclonal antibody (MAb) was applied to a double-antibody sandwich ELISA (DAS ELISA) for highly sensitive detection of cucumber mosaic virus. Alkaline phosphatase-labeled MAb with high activity was used to raise the ratio of spocific-and nonspecific-adsorption of conjugate (S/N ratio) on polyclonal antibody coated solid phase. Also the assay with a fluorogenic substrace or an enzyme-amplified assay which is highly sensitive for measurment of activity was employed to increase the sensitivity of the assays. This ELISA systems proved to be more sensitive than the conventional assay using ρ-nitorophenyl phosphate, and could detect 100 pg/ml of purified virus.
Abstract Alternative
DAS ELISAによるキュウリモザイクウィルスの超高感度検出を試みた。コ-テング抗体(補足抗体)としてのウサギのポリクロナル抗体を、検出抗体(酸素標識抗体)として高活性のMAbコンジュゲートを用いることにより固相への特異的吸着量と非特異的吸着量の比(S/N比)を大きくするとともに、酵素活性の高感度測定に4-methylumbelliferyl phosphateを用いる蛍光法あるいは酵素サイクリング法を用いた。両法はρ-nitrophenyl posphateを用いる比色法よりもウイルスの検出感度が高く、100pg/mlの精製ウイルスを検出することができた。
Keywords
Cucumber mosaic virus
Monoclonal antibody
DAS ELISA
Fluorogenic substrate
Enzyme-amplified assay
Published Date
1993
Publication Title
岡山大学資源生物科学研究所報告
Publication Title Alternative
Bulletin of the Research Institute for Bioresources, Okayama University
Volume
volume1
Issue
issue2
Publisher
岡山大学資源生物科学研究所
Publisher Alternative
Research Institute for Bioresources, Okayama University
Start Page
167
End Page
174
ISSN
0916-930X
NCID
AN10381600
Content Type
Departmental Bulletin Paper
language
日本語
File Version
publisher
Refereed
False