The Search For Inhibitors Of Gram-negative Bacteria’s Metallo-β-lactamase In A Model System | 105741
Journal of AIDS & Clinical Research
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Aim: The study aims to assess the possibility of clodronic acid to inhibit the standard reagent of P. aeruginosa Metallo-β-
Lactamase (MβL)/Sigma, USA/ and to prevent the acquisition of resistance by Gram-negative bacteria that were previously
susceptible to carbapenems.
Method: The final load of sensitive to Meropenem (MIC 2 ?g/ml) reference strains of P. aeruginosa ATCC 27853, A. baumannii
ATCC BAA-747, K. pneumoniae ATCC 70603 was 5??104 CFU in 200 ?l, exposure 24 hours. Checkerboard array was used to
determine the enzyme activity against Meropenem (MIC 2-512 ?g/ml) as well as to evaluate the possibility of clodronic acid
sub-bactericidal doses to inhibit the standard reagent of P. aeruginosa MβL and to prevent the acquisition of resistance to
carbapenems by previously sensitive strains. Suppression of the MβL by clodronic acid and as a result, the lack of growth of the
test strains were confirmed in microcells, as well as on the EL??800 reader (Bio-Tek Instruments Inc., USA).
Result: A dose-dependent effect of acquiring resistance to Meropenem in Gram-negative bacteria (increasing MIC) in the
presence of various amounts of the standard reagent of the enzyme ??βL at an exposure of 24 hours was revealed. At the
same time, P. aeruginosa M?L reagent leads to the emergence of resistance in other types of Gram-negative microorganisms
previously sensitive to carbapenems. Sub-bactericidal doses of clodronic acid do not affect the growth of intact sensitive test
strains, but at the same time its ½ MIC and ¼ MIC completely inhibit the activity of the standard P. aeruginosa MβL reagent
and prevent the acquisition of resistance of previously sensitive to carbapenems Gram-negative bacteria.
Conclusion: Using this method will simplify the search for perspective inhibitors of MβL.
Anna Afinogenova is a Pharmacist, Microbiologist and the Doctor of Biological Science since 2011, specialized in Clinical Microbiology. She is the Head of Laboratory Centre of Saint Petersburg Pasteur Institute and Professor of Saint Petersburg State University. She has published more than 30 papers in profile journals and has 17 patents to her credit. She is the Member of European Wound Management Association since 2010.