Semenov V.M., Zhiltsov I.V., Dmitrachenko T.I., Zenkova S.K., Skvortsova V.V., Kubrakov K.M.,Veremey I.S.
Detection of beta-lactamases production by bacteria with the use of test system «BioLactam»
Abstract.
We have designed the test system «BioLactam» which is capable of registering the level of beta-lactamase activity in biological substrates. In the present study we have evaluated the fundamental possibility of determining beta-lactamase activity of bacterial suspensions using the above-mentioned test system; also we have performed the analysis of the conformance of the obtained results to the data of the routine methods of bacteriological examination. It has turned out that test system «BioLactam» may be successfully used for qualitative and quantitative evaluation of beta-lactamase activity of bacterial suspensions, and the results of testing are in good concordance with those got by the disk diffusion method. By means of test system «BioLactam» we can determine the following things with the high level of reliability: 1) production of clinically significant quantities of beta-lactamases if the level of beta-lactamase activity of bacterial suspension is ≥14,2%; 2) resistance to inhibitor-protected beta-lactams if the level of beta-lactamase activity of bacterial suspension makes up ≥26,5%; 3) resistance to cephalosporins of the 3rd generation if the level of beta-lactamase activity is ≥81,2%; 4) if used together with the disk diffusion method the presence of non-enzymatic mechanisms of resistance to beta-lactam antibiotics as well.
Key words: test system «BioLactam», beta-lactamase activity, bacterial suspensions, antibacterial therapy, disk diffusion method.
References
1. Pérez-Llarena FJ, Bou G. Beta-lactamase inhibitors: the story so far. Curr Med Chem. 2009;16(28):3740-65.
2. Abeylath SC, Turos E. Drug delivery approaches to overcome bacterial resistance to beta-lactam antibiotics. Expert Opin Drug Deliv. 2008 Sep;5(9):931-49.
3. Rolinson GN. Evolution of beta-lactamase inhibitors. Rev Infect Dis. 1991 Jul-Aug;13(9):727-32.
4. Sawai T. Mechanisms of bacterial resistance to beta-lactams by beta-lactamases. Nihon Rinsho. 1997 May;55(5):1225-30.
5. Trehan I, Beadle BM, Shoichet BK. Inhibition of AmpC beta-lactamase through a destabilizing interaction in the active site. Biochemistry. 2001 Jul;40(27):7992-9.
6. National Committee for Clinical Laboratory Standards. Performance standards for antimicrobial susceptibility testing; 10th informational supplement. NCCLS Document M100-S9. 1999. р. 141-156.
7. Reshedko GK, Ctetsiuk OU. Osobennosti opredeleniia chuvstvitel'nosti mikroorganizmov disko-diffuzionnym metodom [Features of determination of sensitivity of microorganisms of the disco - a diffusive method]. Klinicheskaia mikro-biologiia i antimikrobnaia khimioterapiia. 2001;3(4):348-54.
8. Vandepitte J, Engbaek K, Piot P, Heuck CC. Basic laboratory procedures in clinical bacteriology. Geneva: World Health Organization (WHO); 1991. 128 р.
9. Pokrovskii VI, red. Golubeva VI, Kilesso VA, Kiseleva BS. Enterobakterii [Enterobakteriya]: ruk. dlia vrachei. Moscow, RF: Meditsina; 1985. 320 р.
10. Desgrandchamps D. Antibiotics 1992: mechanism of resistance and its clinical relevance. Schweiz Med Wochenschr. 1992 Feb;122(8):247-56.
11. Livermore DM. Mechanisms of resistance to beta-lactam antibiotics. Scand J Infect Dis Suppl. 1991;78:7-16.
12. Medeiros AA, Hedges RW, Jacoby GA. Spread of a «Pseudomonas-specific» beta-lactamase to plasmids of enterobacteria. J Bacteriol. 1982 Feb;149(2):700–7.
13. Michálková-Papajová D, Bujdáková H, Klimácková J, Moravcíková M, Kettner M. Occurrence and mechanisms of resistance to beta-lactam antibiotics in clinically important species of Enterobacter. Epidemiol Mikrobiol Imunol. 2001 Aug;50(3):121-30.
14. Okamoto R, Nakano R. AmpC beta-lactamases producing bacteria. Nihon Rinsho. 2001 Apr;59(4):707-11.
15. Philippon A, Arlet G. Beta-lactamases of Gram negative bacteria: never-ending clockwork. Ann Biol Clin. 2006 Jan-Feb;64(1):37-51.
16. Casalino M, Nicoletti M, Salvia A, Colonna B, Pazzani C, Calconi A, Mohamud KA, Maimone F. Characterization of endemic Shigella flexneri strains in Somalia: antimicrobial resistance, plasmid profiles, and serotype correlation. J Clin Microbiol. 1994 May;32(5):1179-83.
17. O'Callaghan CH, Morris A, Kirby SM, Shingler AH. Novel method for detection of b-lactamases by using a chromogenic cephalosporin substrate. Antimicrob Agents Chemother. 1972 Apr;1(4):283-8.
