US 12,435,352 B2
Mass spectrometric measurement of β-lactamase resistances
Markus Kostrzewa, Lilienthal (DE); Karsten Michelmann, Bremen (DE); and Katrin Sparbier, Bremen (DE)
Assigned to Bruker Daltonics GmbH & Co. KG, Bremen (DE)
Appl. No. 13/582,372
Filed by Markus Kostrzewa, Lilienthal (DE); Karsten Michelmann, Bremen (DE); and Katrin Sparbier, Bremen (DE)
PCT Filed Jun. 10, 2011, PCT No. PCT/EP2011/059670
§ 371(c)(1), (2), (4) Date Dec. 26, 2012,
PCT Pub. No. WO2011/154517, PCT Pub. Date Dec. 15, 2011.
Claims priority of application No. 10 2010 023 452.4 (DE), filed on Jun. 11, 2010.
Prior Publication US 2013/0095511 A1, Apr. 18, 2013
Int. Cl. C12Q 1/18 (2006.01); C12Q 1/34 (2006.01); G01N 33/68 (2006.01)
CPC C12Q 1/18 (2013.01) [C12Q 1/34 (2013.01); G01N 33/6851 (2013.01); G01N 2333/986 (2013.01); G01N 2800/26 (2013.01); G01N 2800/44 (2013.01)] 11 Claims
OG exemplary drawing
 
1. A method for the determination of a β-lactam resistance of microbes based on the production of β-lactamases by the microbes, the method comprising:
obtaining the microbes from a colony cultivated on a nutrient medium or from a blood culture;
combining the microbes with a β-lactam antibiotic covalently bonded to a 6-His tag in a solution, wherein β-lactamases of the microbes may cause hydrolytic breakdown of the β-lactam antibiotic;
extracting remaining 3-lactam antibiotic and/or its breakdown product from the solution via a reversible bond of the 6-His tag to an extracting partner;
partly applying the extracted remaining-lactam antibiotic and/or breakdown product to a mass spectrometric support; and
measuring an enzymatic breakdown of the β-lactam antibiotic by the β-lactamases of the microbes mass spectrometrically by acquiring a mass spectrum of the extracted remaining β-lactam antibiotic and/or breakdown product using a time of flight mass spectrometer with ionization by matrix-assisted laser desorption, wherein a matrix used is α-cyano-4-hydroxycinnamic acid.