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Detection of bacteria from biological mixtures using immunomagnetic separation combined with matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry
Author(s) -
Madonna Angelo J.,
Basile Franco,
Furlong Ed,
Voorhees Kent J.
Publication year - 2001
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.344
Subject(s) - chemistry , chromatography , mass spectrometry , immunomagnetic separation , matrix (chemical analysis) , bacteria , desorption , matrix assisted laser desorption/ionization , microorganism , analytical chemistry (journal) , adsorption , biology , genetics , organic chemistry
A rapid method for identifying specific bacteria from complex biological mixtures using immunomagnetic separation coupled to matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry has been developed. The technique employs commercially available magnetic beads coated with polycolonal antibodies raised against specific bacteria and whole cell analysis by MALDI‐MS. A suspension of a bacterial mixture is mixed with the immunomagnetic beads specific for the target microorganism. After a short incubation period (20 mins) the bacteria captured by the beads are washed, resuspended in deionized H 2 O and directly applied onto a MALDI probe. Liquid suspensions containing bacterial mixtures can be screened within 1 h total analysis time. Positive tests result in the production of a fingerprint mass spectrum primarily consisting of protein biomarkers characteristic of the targeted microorganism. Using this procedure, Salmonella choleraesuis was isolated and detected from standard bacterial mixtures and spiked samples of river water, human urine, and chicken blood. Copyright © 2001 John Wiley & Sons, Ltd.