Discrimination of clostridium species using a magnetic bead based hybridization assay

Susanne Pahlow; Barbara Seise; Sibyll Pollok; Christian Seyboldt; Karina Weber; Jürgen Popp

doi:10.1117/12.2051340

8 May 2014 Discrimination of clostridium species using a magnetic bead based hybridization assay

Susanne Pahlow, Barbara Seise, Sibyll Pollok, Christian Seyboldt, Karina Weber, Jürgen Popp

Proceedings Volume 9129, Biophotonics: Photonic Solutions for Better Health Care IV; 91292J (2014) https://doi.org/10.1117/12.2051340
Event: SPIE Photonics Europe, 2014, Brussels, Belgium

Abstract

Clostridium chauvoei is the causative agent of blackleg, which is an endogenous bacterial infection. Mainly cattle and other ruminants are affected. The symptoms of blackleg are very similar to those of malignant edema, an infection caused by Clostridium septicum. [1, 2] Therefore a reliable differentiation of Clostridium chauvoei from other Clostridium species is required. Traditional microbiological detection methods are time consuming and laborious. Additionally, the unique identification is hindered by the overgrowing tendency of swarming Clostridium septicum colonies when both species are present. [1, 3, 4] Thus, there is a crucial need to improve and simplify the specific detection of Clostridium chauvoei and Clostridium septicum. Here we present an easy and fast Clostridium species discrimination method combining magnetic beads and fluorescence spectroscopy. Functionalized magnetic particles exhibit plentiful advantages, like their simple manipulation in combination with a large binding capacity of biomolecules. A specific region of the pathogenic DNA is amplified and labelled with biotin by polymerase chain reaction (PCR). These PCR products were then immobilized on magnetic beads exploiting the strong biotin-streptavidin interaction. The specific detection of different Clostridium species is achieved by using fluorescence dye labeled probe DNA for the hybridization with the immobilized PCR products. Finally, the samples were investigated by fluorescence spectroscopy. [5]

Citation Download Citation

Susanne Pahlow, Barbara Seise, Sibyll Pollok, Christian Seyboldt, Karina Weber, and Jürgen Popp "Discrimination of clostridium species using a magnetic bead based hybridization assay", Proc. SPIE 9129, Biophotonics: Photonic Solutions for Better Health Care IV, 91292J (8 May 2014); https://doi.org/10.1117/12.2051340

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