IODINE TEST FOR EXPRESS ESTIMATION OF BETA-LACTAMASE ACTIVITY IN ISOLATES OF STAPHYLOCOCCUS AUREUS

DOI: https://doi.org/10.29296/25877313-2020-07-06
Issue: 
7
Year: 
2020

A.N. Vaganova Ph.D. (Biol), Research Scientist, Laboratory of Molecular Biological Technologies, Institut Pasteur in Saint-Petersburg for Research in Epidemiology and Microbiology, Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing (Saint Petersburg) E-mail: van.inprogress@gmail.com M.V. Micheeva Laboratory Assistant, Laboratory of Molecular Biological Technologies, Institut Pasteur in Saint-Petersburg for Research in Epidemiology and Microbiology, Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing (Saint Petersburg) E.V. Nesterova Bacteriologist, Clinical Diagnostic Laboratory, City Skin and Veneral Diseases Dispensary (Saint Petersburg) N.N. Trofimova Bacteriologist, Clinical Diagnostic Laboratory, City Skin and Veneral Diseases Dispensary(Saint Petersburg) I.V. Litvinenko Head of Clinical Diagnostic Laboratory, City Skin and Veneral Diseases Dispensary(Saint Petersburg) Y.G. Petunova Ph.D. (Med.), City Skin and Veneral Diseases Dispensary(Saint Petersburg) V.N. Verbov Ph.D. (Chem.), Institut Pasteur in Saint-Petersburg for Research in Epidemiology and Microbiology, Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing (Saint Petersburg)

Background. Beta-lactamases are widely spreadin methicillin-susceptible Staphylococcus aureus (MSSA) isolates and confer this pathogen spenicillin-resistance. At the same time 50% of isolates that are penicillin-susceptible in disk diffusion test are beta-lactamase producers. Antibiotic resistance evaluation is time-consumin gand prolongs the process of adequate treatment prescribing. So the introduction of express-tests for antibiotic resistance evaluation is an actual problem. The aim of this study. The estimation of iodine test sensitivity for penicillin susceptibility and potential beta-lactamase activity in staphylococci (i.e. isolates with beta-lactamase operon in their genomes). Materials and methods. Beta-lactamase activity of 81 MSSA clinical isolates was evaluated by iodine test with preliminary stimulation of beta-lactamase production and without it. To estimate the sensitivity and specificity of this method for identification of penicillin-resistant isolates and po-tential beta-lactamase producers the results of the iodine test were compared with the data about beta-lactamase operon presence in isolates ge-nomes recived in real-time PCR and statements of penicillin-resistance in disk diffusion test. Results. Iodine test was highly sensitive for identification penicillin-susceptible MSSA isolates. The sensitivity of iodine test for penicillin-resistant is olates was 61% if the test was formed without preliminary stimulation or 86% if the test was performed after stimulation. In the same time iodine test was low-sensitive for isolates that contain beta-lactamase operon in their genomes identification, the iodine test sensitivity in this case was less than 63%. So iodine test without preliminary stimulation maybe regarded as possible screening method for penicillin-susceptible MSSA identification immediately after pathogen isolation, that will be helpful to decrease the time-loss for decision making in antibiotic treatment choose. Conclusion. Despite the low sensitivity of iodine test for identification of beta-lactamase operon containing MSSA, this method is attractive for fast identification of penicillin-resistance in S. aureus.

Keywords: 
Staphylococcus aureus
beta-lactamase
antibiotic resistance
diagnosis

References: 
  1. Song K.H., Jung S.-I., Lee S., Park S., Kim E.S:,Park K.-H., Park W.B., Choe P.G., Kim Y.K., Kwak Y.G., Kim Y.S., Jang H.C., Kiem S., Kim H.I., Kim H.B. Inoculum effect of methicillin-susceptible Staphylococcus aureus against broad-spectrum beta-lactamantibiotics. Eur. J. Clin. Microbiol. Infect. Dis. 2019; 38(1):67-74.
  2. Olsen J.E., Christensen H., Aarestrup F.M. Diversity and evolution of blaZ from Staphylococcus aureus and coagulase-negative staphylococci. J Antimicrob. Chemother. 2006; 57(3): 450–460.
  3. Pereira L.A., Harnett G.B., Hodge M.M., Cattell J.A., Speers D.J. Real-time PCR assay for detection of blaZ genes in Staphylococcus aureus clinical isolates. J. Clin. Microbiol. 2014; 52(4): 1259–1261.
  4. Ruegg P.L., L Oliveira L., Jin W., Okwumabua O. Phenotypic antimicrobial susceptibility and occurrence of selected resistance genes in gram-positive mastitis pathogens isolated from wisconsin dairy cows. J. Dairy Sci. 2015; 98(7):4521–4534.
  5. Vento T.J., Calvano T.P., Cole D.W., Mende K., Rini E.A., Tully C.C., Landrum M.L., Zera W., Guymon C.H., Yu X., Beckius M.L., Cheatle K.A., Murray C.K. Staphylococcus aureus colonization of healthy military service members in the United States and Afghanistan. BMC Infect. Dis. 2013; 13:325.
  6. Kaase M., Lenga S., Friedrich S., Szabados F., Sakinc T., Kleine B., Gatermann S.G. Comparison of phenotypic methods for penicillinase detection in Staphylococcus aureus. Clin. Microbiol. Infect. 2008; 14(6):614–616.
  7. Jagad B.P., Vakanee A. Comparison of three different methods to detect the production of β-lactamase enzyme by Staphylococci. International Journal of Biomedical and Advance Research. 2017; 8(1):01–03.
  8. Al-Hammadi A.T.Y. Comparative study of two beta-lactamase test methods in five different microorganisms isolated from Azadi Hospital in Kirkuk. Iraq International Journal of Scientific and Research Publications. 2016; 6(10):2250–3153.
  9. Bidya S., Suman R.S. Comparative Study of Three β Lactamase Test Methods in Staphylococcus aureus Isolated from Two Nepalese Hospitals. Open Journal of Clinical Diagnostics. 2014; 4:47–52.
  10. EUCAST. 2015. The European Committee on Antimicrobial Susceptibility Testing. Breakpoint tables for interpretation of MICs and zone diameters. Version 9.0, valid from 2019-01-01. http://www.eucast.org. Accessed 30/11/2019.