NDLI: <i>Clostridium</i> spp. discrimination with a simple bead-based fluorescence assay

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Analytical Methods Vol - 6 Issue - 8

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Clostridium spp. discrimination with a simple bead-based fluorescence assay

Content Provider	Royal Society of Chemistry (RSC)
Author	Seyboldt, Christian Neubauer, Heinrich Popp, Jürgen Klapper, Martin Weber, Karina Pollok, Sibyll Seise, Barbara Pahlow, Susanne
Copyright Year	2014
Abstract	C. chauvoei is the causative agent of blackleg, an endogenous bacterial infection which usually affects cattle and other ruminants. Due to the fact that the symptoms of this severe disease are very similar to the phenotype caused by an infection with C. septicum, a reliable differentiation of C. chauvoei from other Clostridium spp. is mandatory. Traditional microbiological detection methods are time consuming and the proper specification is hampered by the overgrowing tendency of swarming C. septicum colonies when both species are in the clinical sample. Thus, there is an urgent need to improve and simplify the specific detection of C. chauvoei and C. septicum. We report an easy and fast Clostridium spp. discrimination method via a magnetic bead-based fluorescence assay. To that end, the target DNA was amplified using 16S-23S rDNA spacer region specific primers. These PCR products were employed to generate single-stranded capture probe DNA, which was immobilized on magnetic beads. Functionalized magnetic particles exhibit numerous advantages, like their simple manipulation in combination with a huge binding capacity of biomolecules and make therefore excellent biosensors. In this context, the discrimination between C. chauvoei and C. septicum was realized by means of hybridization with complementary detection probe DNA. Finally, fluorescence spectroscopy allowed the signal readout. With this approach a precise discrimination between C. chauvoei, C. septicum and C. carnis was accomplished.
Starting Page	2943
Ending Page	2949
Page Count	7
File Format	HTM / HTML PDF
ISSN	17599660
Volume Number	6
Issue Number	9
Journal	Analytical Methods
DOI	10.1039/c3ay41888j
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Strikebreaker Clostridium Polymerase chain reaction Spectroscopy Phenotype 16S ribosomal RNA DNA Fluorescence Sepsis 23S ribosomal RNA
Content Type	Text
Resource Type	Article
Subject	Analytical Chemistry Engineering Chemical Engineering

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Clostridium spp. discrimination with a simple bead-based fluorescence assay

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Clostridium spp. discrimination with a simple bead-based fluorescence assay