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基于干试剂的 PCR 作为一种新型工具,用于快速检测梭菌属。

Dry-reagent-based PCR as a novel tool for the rapid detection of Clostridium spp.

机构信息

Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich-Schiller-University Jena, Helmholtzweg 4, 07743 Jena, Germany.

Institute of Photonic Technology Jena, Albert-Einstein-Straße 9, 07745 Jena, Germany.

出版信息

J Med Microbiol. 2013 Oct;62(Pt 10):1588-1591. doi: 10.1099/jmm.0.060061-0. Epub 2013 Jul 31.

Abstract

Improved conventional PCR techniques are required for the rapid on-site detection of human and animal diseases. In this context, a PCR method using dry-stored reagents intended for the detection of Clostridium spp. is presented. Basic PCR reagents (BSA, PCR buffer, MgCl₂ and primers), which were dried on polyolefin matrices, showed stability at ambient temperatures for up to 10 months without any loss of functionality. An outstanding advantage of our amelioration is the elimination of PCR process errors caused by the improper storage and handling of liquid reagents. Moreover, our PCR-based amplification can be performed in less than 30 min, saving time compared with conventional detection methods. Thus, dry-reagent-based PCR is implementable in a suitcase-like modular device for the rapid on-site detection of microbial pathogens such as blackleg of ruminants caused by Clostridium chauvoei.

摘要

需要改进的常规聚合酶链反应技术用于快速现场检测人类和动物疾病。在这种情况下,提出了一种使用干燥储存试剂检测梭菌属的 PCR 方法。基本的 PCR 试剂(BSA、PCR 缓冲液、MgCl₂和引物)在聚烯烃基质上干燥,在环境温度下稳定长达 10 个月,没有任何功能丧失。我们改进的一个突出优点是消除了由于液体试剂储存和处理不当而导致的 PCR 过程错误。此外,我们基于 PCR 的扩增可以在不到 30 分钟内完成,与传统检测方法相比节省了时间。因此,基于干燥试剂的 PCR 可在手提箱式模块化设备中实施,用于快速现场检测由 C. chauvoei 引起的反刍动物坏疽等微生物病原体。

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