Chapter title |
Detection of Yersinia pestis in Complex Matrices by Intact Cell Immunocapture and Targeted Mass Spectrometry
|
---|---|
Chapter number | 7 |
Book title |
Microbial Toxins
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6958-6_7 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6956-2, 978-1-4939-6958-6
|
Authors |
Jérôme Chenau, François Fenaille, Stéphanie Simon, Sofia Filali, Hervé Volland, Christophe Junot, Elisabeth Carniel, François Becher |
Editors |
Otto Holst |
Abstract |
We describe an immunoaffinity-liquid chromatography-tandem mass spectrometry (immuno-LC-MS/MS) protocol for the direct (i.e., without prior culture), sensitive and specific detection of Yersinia pestis in complex matrices. Immunoaffinity enables isolation and concentration of intact bacterial cells from food and environmental samples. After protein extraction and digestion, suitable proteotypic peptides corresponding to three Y. pestis-specific protein markers (murine toxine, plasminogen activator and pesticin) are monitored by targeted LC-MS/MS using the selected reaction monitoring (SRM) mode. This immuno-LC-MS/MS assay has a limit of detection of 2 × 10(4) CFU/mL in milk or tap water, and 4.5 × 10(5) CFU in 10 mg of soil. |
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