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The consumption of counterfeit ART drugs has resulted in the death of many patients. Thus,evaluating the quality of ART drugs is needed. There are several methods for quantitating the ART content in tablets,the most common being a high-performance liquid chromatography. However,that method is hampered by the need for expensive equipment and a rather time-consuming process of extraction. By contrast,enzyme-linked immunosorbent assays (ELISAs) are faster and much less expensive,and they require less sample preparation than the above method. The objective of the present study was to establish a simple and specific direct competitive ELISA for the determination of ART concentrations using an anti-ART polyclonal antibody (pAb). Results: Anti-ART pAb was raised in mice,and ART-horseradish peroxidase (HRP) conjugate was produced. A direct competitive ELISA was performed by simultaneously incubating ART and the ART-HRP conjugate with the anti-ART pAb over a second antibody. Subsequently,the enzyme activity of the remaining ART-HRP conjugate was measured. The intra-and inter-assay coefficients of variation of the ELISA were less than 10 % in the range of 0.3 to 30 ng/ml with a detection limit of 0.1 ng/ml. The cross-reactivities of the anti-ART pAb with ARM and dihydroartemisinin were 0.12 and 0.04 %,respectively,and those with other antimalarial drugs were negligible. Furthermore,the recovery of 10 or 50 ng/ml ART added to the drug tablet solutions containing an expected amount of 10 ng/ml was estimated by the ELISA. The recovery of the ART amount ranged between 98 and 106 %,with coefficient variations of less than 7.0 %. Conclusions: The present ELISA is a simple and specific method for the determination of ART concentrations. 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Development of a simple and specific direct competitive ELISA for the determination of artesunate using an anti-artesunate polyclonal antiserum
http://hdl.handle.net/10069/37352
http://hdl.handle.net/10069/37352efea92bc-6b55-4d66-8938-7ea333797bc9
名前 / ファイル | ライセンス | アクション |
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TMH44_37.pdf (551.0 kB)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2017-05-25 | |||||
タイトル | ||||||
タイトル | Development of a simple and specific direct competitive ELISA for the determination of artesunate using an anti-artesunate polyclonal antiserum | |||||
言語 | ||||||
言語 | eng | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Artesunate | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | ELISA | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Malaria | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Polyclonal antibody | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
著者 |
Mitsui, Yoshinori
× Mitsui, Yoshinori |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Background: Since artesunate (ART) became a vital component of artemisinin (ARM)-based combination therapies for the treatment for malaria,counterfeit ART drugs have spread in regions of Southeast Asia and Africa. The consumption of counterfeit ART drugs has resulted in the death of many patients. Thus,evaluating the quality of ART drugs is needed. There are several methods for quantitating the ART content in tablets,the most common being a high-performance liquid chromatography. However,that method is hampered by the need for expensive equipment and a rather time-consuming process of extraction. By contrast,enzyme-linked immunosorbent assays (ELISAs) are faster and much less expensive,and they require less sample preparation than the above method. The objective of the present study was to establish a simple and specific direct competitive ELISA for the determination of ART concentrations using an anti-ART polyclonal antibody (pAb). Results: Anti-ART pAb was raised in mice,and ART-horseradish peroxidase (HRP) conjugate was produced. A direct competitive ELISA was performed by simultaneously incubating ART and the ART-HRP conjugate with the anti-ART pAb over a second antibody. Subsequently,the enzyme activity of the remaining ART-HRP conjugate was measured. The intra-and inter-assay coefficients of variation of the ELISA were less than 10 % in the range of 0.3 to 30 ng/ml with a detection limit of 0.1 ng/ml. The cross-reactivities of the anti-ART pAb with ARM and dihydroartemisinin were 0.12 and 0.04 %,respectively,and those with other antimalarial drugs were negligible. Furthermore,the recovery of 10 or 50 ng/ml ART added to the drug tablet solutions containing an expected amount of 10 ng/ml was estimated by the ELISA. The recovery of the ART amount ranged between 98 and 106 %,with coefficient variations of less than 7.0 %. Conclusions: The present ELISA is a simple and specific method for the determination of ART concentrations. Thus,this ELISA can be used to identify ART counterfeits and substandard drugs and to quantify the ART drugs. | |||||
書誌情報 |
Tropical Medicine and Health 巻 44, p. 37, 発行日 2016-11-21 |
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出版者 | ||||||
出版者 | BioMed Central Ltd. | |||||
EISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 13494147 | |||||
DOI | ||||||
関連タイプ | isIdenticalTo | |||||
識別子タイプ | DOI | |||||
関連識別子 | 10.1186/s41182-016-0037-2 | |||||
権利 | ||||||
権利情報 | c The Author(s) 2016. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. | |||||
著者版フラグ | ||||||
出版タイプ | VoR | |||||
出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||
引用 | ||||||
内容記述タイプ | Other | |||||
内容記述 | Tropical Medicine and Health, 44, 37; 2016 |