Rapid detection of porins by matrix-assisted laser desorption/ionization-time of flight mass spectrometry

Rapid detection of porins by matrix-assisted laser desorption/ionization-time of flight mass spectrometry

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The rapid and cost-efficient determination of carbapenem resistance is an important prerequisite for the choice of an adequate antibiotic therapy.A MALDI-TOF MS-based assay was set up to detect porins in the current study.A loss of the components of porin alone such as OmpK35/OmpK36 or together with the production of carbapenemases will augment the carbapenem resistance.

Ten strains of E.coli voyage et cie discount code and eight strains of K.pneumoniae were conducted for both SDS-PAGE and MALDI-TOF MS analysis.

MALDI-TOF/TOF MS analysis was then performed to verify the corrospondence of proteins between SDS-PAGE and MALDI-TOF MS.The results indicated that the mass spectrum of ca.35,000-m/z, 37,000-m/z and 38,000-m/z peaks of E.

coli ATCC 25922 corresponded to OmpA, OmpC and OmpF with molecular weight of approximately ca.38 kDa, 40 kDa and 41 kDa in SDS-PAGE gel, respectively.The band of OmpC and OmpF porins were unable to be distinguished by SDS-PAGE, whereas blackmores ache relief focus review it was easy to be differentiated by MALDI-TOF MS.

As for K.pneumoniae isolates, the mass spectrum of ca.36,000-m/z and 38,600-m/z peaks was observed corresponding to OmpA and OmpK36 with molecular weight of approximately ca.

40 kDa and 42 kDa in SDS-PAGE gel, respectively.Porin OmpK35 was not observed in the current SDS-PAGE, while a 37,000-m/z peak was found in K.pneumoniae ATCC 13883 and carbapenem-susceptible strains by MALDI-TOF MS which was presumed to be the characteristic peak of the OmpK35 porin.

Compared with SDS-PAGE, MALDI-TOF MS is able to rapidly identify the porin-deficient strains within half an hour with better sensitivity, less cost, and is easier to operate and has less interference.