Rapid detection of virulent protease secreted by Vibrio anguillarum by dot enzyme-linked immunosorbent assay
文献类型：期刊论文
| 作者 | Zhang, ZD; Zhang, PJ; Mo, ZL; Wang, CL; Yu, Y
|
| 刊名 | ACTA OCEANOLOGICA SINICA
 |
| 出版日期 | 2005 |
| 卷号 | 24期号:4页码:155-161 |
| 关键词 | Vibrio Anguillarum Extracellular Products Protease Dot-elisa Indirect Elisa Western Blot |
| ISSN号 | 0253-505X |
| 文献子类 | Article |
| 英文摘要 | Dot enzyme-linked immunosorbent assay (dot-ELISA), indirect ELISA and Western blot were performed to detect the virulent protease secreted by Vibrio anguillarum which was isolated from the diseased left-eyed flounder, Paralichthys olivaceous. Sensitivity results showed that dot-ELISA is a more sensitive, rapid and simple technique for the protease detection. The minimal detectable amount of protease is about 7 pg in the dot-ELISA test, while 7.8 ng in the indirect ELISA and 6.25 ng in the Western blot respectively. Protease could be detected 2 h after incubation of V. anguillarum in the 2216E liquid medium but enzyme activity was very low at that period. From 6 to 12 h, the amount and enzyme activity of protease increased markedly and reached maximum at stationary phase. Analysis of serum samples periodically collected from the infected flounders showed that after 2 h of infection by V. anguillarum, the pathogenic bacteria could be detected in the blood of the infected flounders but no protease was found. It was 5 similar to 6 h after infection that the protease was detected in blood and then the amount increased as infection advanced. Quantitative detection of protease either incubation in the medium or from the blood of infected flounders could be accomplished in virtue of positive controls of quantificational protease standards ("marker") so that the alterations of protease secretion both in vitro and in vivo could be understood generally. In addition, the indirect ELISA and dot-ELISA were also performed to detect V. anguillarum cells. Results indicated that the sensitivity of indirect ELISA to bacteria cells is higher than that of the dot-ELISA, and that the minimal detectable amount is approximately 10(4) cell/mL in the indirect ELISA, while 10(5) cell/mL in the dot-ELISA.; Dot enzyme-linked immunosorbent assay (dot-ELISA), indirect ELISA and Western blot were performed to detect the virulent protease secreted by Vibrio anguillarum which was isolated from the diseased left-eyed flounder, Paralichthys olivaceous. Sensitivity results showed that dot-ELISA is a more sensitive, rapid and simple technique for the protease detection. The minimal detectable amount of protease is about 7 pg in the dot-ELISA test, while 7.8 ng in the indirect ELISA and 6.25 ng in the Western blot respectively. Protease could be detected 2 h after incubation of V. anguillarum in the 2216E liquid medium but enzyme activity was very low at that period. From 6 to 12 h, the amount and enzyme activity of protease increased markedly and reached maximum at stationary phase. Analysis of serum samples periodically collected from the infected flounders showed that after 2 h of infection by V. anguillarum, the pathogenic bacteria could be detected in the blood of the infected flounders but no protease was found. It was 5 similar to 6 h after infection that the protease was detected in blood and then the amount increased as infection advanced. Quantitative detection of protease either incubation in the medium or from the blood of infected flounders could be accomplished in virtue of positive controls of quantificational protease standards ("marker") so that the alterations of protease secretion both in vitro and in vivo could be understood generally. In addition, the indirect ELISA and dot-ELISA were also performed to detect V. anguillarum cells. Results indicated that the sensitivity of indirect ELISA to bacteria cells is higher than that of the dot-ELISA, and that the minimal detectable amount is approximately 10(4) cell/mL in the indirect ELISA, while 10(5) cell/mL in the dot-ELISA. |
| 学科主题 | Oceanography |
| 语种 | 英语 |
| WOS记录号 | WOS:000231165800018 |
| 公开日期 | 2010-12-22 |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/3167]  |
| 专题 | 海洋研究所_实验海洋生物学重点实验室 |
| 作者单位 | 1.Chinese Acad Sci, Inst Oceanol, Key lab Expt Marine Biol, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhang, ZD,Zhang, PJ,Mo, ZL,et al. Rapid detection of virulent protease secreted by Vibrio anguillarum by dot enzyme-linked immunosorbent assay[J]. ACTA OCEANOLOGICA SINICA,2005,24(4):155-161. |
| APA | Zhang, ZD,Zhang, PJ,Mo, ZL,Wang, CL,&Yu, Y.(2005).Rapid detection of virulent protease secreted by Vibrio anguillarum by dot enzyme-linked immunosorbent assay.ACTA OCEANOLOGICA SINICA,24(4),155-161. |
| MLA | Zhang, ZD,et al."Rapid detection of virulent protease secreted by Vibrio anguillarum by dot enzyme-linked immunosorbent assay".ACTA OCEANOLOGICA SINICA 24.4(2005):155-161. |
入库方式： OAI收割
来源：海洋研究所
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