Rapid and Sensitive detection of Staphylococcus aureus and Klebsiella pneumonia based on Bacitracin-modified Fe3O4@PDA magnetic beads combined with Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

2021 ◽  
Author(s):  
Feng Qin ◽  
Shujuan Wang ◽  
Mingxia Gao ◽  
Xiangmin Zhang
Keyword(s):  
Mass Spectrometry ◽  
Magnetic Beads ◽  
Laser Desorption ◽  
Klebsiella Pneumonia ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Desorption Ionization ◽  
Flight Mass Spectrometry ◽  
Matrix Assisted Laser ◽  
Matrix Assisted Laser Desorption

The highly effective detection of pathogens in complex biological samples is an attractive and critical topic all the time. Bacitracin is a novel broad-spectrum antimicrobial peptide to enrich bacteria via...

scholarly journals Coupling Immunomagnetic Separation on Magnetic Beads with Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Detection of Staphylococcal Enterotoxin B

2007 ◽  
Vol 73 (21) ◽  
pp. 6945-6952 ◽  
Author(s):  
Gitta Schlosser ◽  
Petr Kačer ◽  
Marek Kuzma ◽  
Zolt�n Szil�gyi ◽  
Alida Sorrentino ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Magnetic Beads ◽  
Laser Desorption ◽  
Molecular Probe ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Desorption Ionization ◽  
Flight Mass Spectrometry ◽  
Matrix Assisted Laser ◽  
Matrix Assisted Laser Desorption

ABSTRACT The growing importance of mass spectrometry for the identification and characterization of bacterial protein toxins is a consequence of the improved sensitivity and specificity of mass spectrometry-based techniques, especially when these techniques are combined with affinity methods. Here we describe a novel method based on the use of immunoaffinity capture and matrix-assisted laser desorption ionization-time of flight mass spectrometry for selective purification and detection of staphylococcal enterotoxin B (SEB). SEB is a potent bacterial protein toxin responsible for food poisoning, as well as a potential biological warfare agent. Unambiguous detection of SEB at low-nanogram levels in complex matrices is thus an important objective. In this work, an affinity molecular probe was prepared by immobilizing anti-SEB antibody on the surface of para-toluene-sulfonyl-functionalized monodisperse magnetic particles and used to selectively isolate SEB. Immobilization and affinity capture procedures were optimized to maximize the density of anti-SEB immunoglobulin G and the amount of captured SEB, respectively, on the surface of magnetic beads. SEB could be detected directly “on beads” by placing the molecular probe on the matrix-assisted laser desorption ionization target plate or, alternatively, “off beads” after its acidic elution. Application of this method to complex biological matrices was demonstrated by selective detection of SEB present in different matrices, such as cultivation media of Staphylococcus aureus strains and raw milk samples.

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