Simultaneous detection of two bacterial pathogens using bacteriophage amplification coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.
The simultaneous analysis of multiple target microorganisms using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) with bacteriophage amplification is discussed. Following infection of target bacteria with specific bacteriophages, proteins contained in the progeny phage are utilized as a secondary biomarker for the target bacterium. Escherichia coli when mixed with MS2 and MPSS-1 phages specific for E. coli and Salmonella spp., respectively, at levels below their corresponding detection limits, produced only the protein (13.7 kDa) characteristic of the MS2. Likewise, Salmonella spp. when mixed with the two phages only produced a protein (13.5 kDa) characteristic of MPSS-1. When the two bacteria and the two phages were mixed together, proteins characteristic of MS2 and MPSS-1 were observed indicating that both bacteriophages had been amplified. Identification of each bacterium was made based on the presence of the secondary bacteriophage biomarkers. No deleterious effects on bacteriophage amplification were observed because of the presence of multiple bacteria or bacteriophages.
Rees
Jon C
JC
Department of Chemistry and Geochemistry, Colorado School of Mines, Golden, CO 80401, USA.
Voorhees
Kent J
KJ
D001435:Bacteriophages / Q000378:metabolism*; D015169:Colony Count, Microbial / Q000379:methods; D004926:Escherichia coli / Q000302:isolation & purification* / Q000821:virology*; D012475:Salmonella / Q000302:isolation & purification* / Q000821:virology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014764:Viral Proteins / Q000032:analysis*
D014764:Viral Proteins