一个基于菌体的平台,用于快速检测蛋白酶的痕迹
Petr Capek1, Killeen S Kirkconnell, Tobin J Dickerson
1Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Journal of the American Chemical Society
|September 4, 2010
概括
我们开发了一种使用菌体检测强大的细菌毒素,如肉毒神经毒素 (BoNT) 和炭致命因子的敏感,廉价的测试方法. 该平台为临床诊断提供基于活动的快速蛋白酶量化.
科学领域:
- 生物技术是生物技术.
- 分析化学 分析化学
- 微生物学 微生物学
背景情况:
- 敏感,快速和廉价的蛋白酶活性测试对于临床诊断至关重要.
- 检测来自Clostridium botulinum和Bacillus anthracis的高强度细菌毒素需要非常敏感.
研究的目的:
- 开发一种廉价和灵敏的测试平台,用于基于活动的蛋白质酶量化.
- 将这个平台应用于检测细菌毒素,特别是肉毒神经毒素 (BoNT) 和炭病致命因子.
主要方法:
- 利用有线细菌作为一个可指数增强的记者系统.
- 基于细菌从固体支物的特定裂变进行测定.
- 通过传染性或定量PCR来量化分离的细菌菌体.
主要成果:
- 已证明检测了甲型和乙型肉毒神经毒素 (BoNT) 的检测.
- 成功检测到炭病致命因子.
- 在优化条件下,BoNT/A的检测极限为2 pM.
结论:
- 开发的基于菌体的测定方法是廉价的,灵敏的,快速的.
- 该平台有效用于基于活动的蛋白酶量化.
- 这种测试显示了临床诊断中危险细菌毒素的敏感检测的前景.
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