Universal extraction method for gastrointestinal pathogens
| Autor: | F. J. Bolton, Rohini Manuel, Xose Couto-Parada, Spencer D. Polley, James J. Gray, Adele V. Lee, John Wain, Fenella D. Halstead, Miren Iturriza-Gomara, Clare Ling, Claire Jenkins, Rachel M. Chalmers, Kristin Elwin, Duncan A. Clark |
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| Rok vydání: | 2013 |
| Předmět: |
Microbiology (medical)
Cycle threshold Time Factors Extraction (chemistry) Cryptosporidium Gastrointestinal pathogens Bacterial Infections DNA General Medicine Biology Real-Time Polymerase Chain Reaction biology.organism_classification Isolation (microbiology) Microbiology Gastroenteritis Specimen Handling Feces Molecular Diagnostic Techniques London Nucleic acid Humans Extraction methods Intestinal Diseases Parasitic Bacteria |
| Zdroj: | Journal of Medical Microbiology. 62:1535-1539 |
| ISSN: | 1473-5644 0022-2615 |
| DOI: | 10.1099/jmm.0.058743-0 |
| Popis: | A universal stool extraction method for recovery of nucleic acids (NAs) from gastrointestinal pathogens was developed to support rapid diagnostics for the London 2012 Olympics. The method involved mechanical disruption (bead beating) of the stools, followed by automated extraction and detection using real-time PCR. This method had been used extensively in the Second Infectious Intestinal Disease Study (IID2) for the isolation of NA from bacteria and parasites (and was effective for the robust recovery of Cryptosporidium spp.) but had not been used for enteric viruses. To ensure this method was universally suitable, panels of samples known to contain target bacteria, viruses or parasites were processed in triplicate using the pre-treatment method routinely used for each target and the new extraction method (bead beating). The extracts were tested using real-time PCR and the cycle threshold values were compared. The results from this study showed that bead beating improved yields for the bacterial and parasitic targets and was suitable for the viral targets. The implementation of this universal method should confer cost- and time-saving benefits and streamline the processes required for the characterization of an array of pathogens from faecal samples. |
| Databáze: | OpenAIRE |
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