×
Home Current Archive Editorial board
News Contact
Review paper

Development of a novel biofilm classification tool and comparative analysis of result interpretation methodologies for the evaluation of biofilm forming capacity of bacteria using tissue culture plate method

By
Monia Avdić Orcid logo ,
Monia Avdić
Contact Monia Avdić

International Burch University, Sarajevo, Bosnia and Herzegovina

Nermin Džuzić ,
Nermin Džuzić

International Burch University, Sarajevo, Bosnia and Herzegovina

Osman Hasanić ,
Osman Hasanić

International Burch University, Sarajevo, Bosnia and Herzegovina

Amel Spahić ,
Amel Spahić

Mistral Technologies, Sarajevo, Bosnia and Herzegovina

Lejla Smajlović Skenderagić ,
Lejla Smajlović Skenderagić

International Burch University, Sarajevo, Bosnia and Herzegovina

Almir Badnjević ,
Almir Badnjević

International Burch University, Sarajevo, Bosnia and Herzegovina

Verlab Ltd., Sarajevo, Bosnia and Herzegovina

Mirsada Hukić
Mirsada Hukić

Institute for Biomedical Diagnostics and Research NALAZ, Sarajevo, Bosnia and Herzegovina

Academy of Sciences and Arts of Bosnia and Herzegovina, Sarajevo, Bosnia and Herzegovina

Abstract

Aim
To develop an online biofilm calculation tool (Biofilm Classifier), which calculates the optical density cut off value and accordingly determines the biofilm forming categories for the tested isolates by standardized formulas, as well as to compare the results obtained by Biofilm Classifier to manual calculations and the use of predefined values.
Methods
The biofilm forming capacity of tested strains was evaluated using tissue culture plate method in 96 well plates, and optical density (OD) value of the formed biofilm was measured on an ELISA Microplate reader at 595 nm on a total of 551 bacterial isolates from clinical specimen.
Results
Comparative analysis indicated that the manual calculation was 100% in accordance with results obtained by the designed software as opposed to the results obtained by use of predefined values for biofilm categorization. When using predefined values compared to manual biofilm categorization for the determination of biofilm positive and biofilm negative strains the specificity was 100%, sensitivity 97.81%, positive predictive value 100%, negative predictive value 96.04% and accuracy 98.57%.
Conclusion
Considering obtained results, the use of the designed online calculator would simplify the interpretation of biofilm forming capacity of bacteria using tissue culture plate method.

References

1.
Ibrišimović M, Ibrišimović M, Mehmedinović N, Hukić M. A novel spectrophotometric assay for the determination of biofilm forming capacity of causative agents of urinary tract infections. IJERT. 2016. p. 1225–30.
2.
Hukić M, Seljmo D, Ramovic A, Ibrišimović M, Dogan S, Hukic J, et al. The effect of lysozyme on reducing biofilms by Staphylococcus aureus, Pseudomonas aeruginosa, and Gardnerella vaginalis: an in vitro examination. Microb Drug Resist. 2018. p. 353–8.
3.
Lewandowski Z, Beyenal H. Fundamentals of Biofilm Research. CRC Press; 2013.
4.
Mcdonnell G, Russell A. Antiseptics and disinfectants: activity, action, and resistance. Clin Microbiol Rev. 2001. p. 227.
5.
O’toole G, Pratt L, Watnick P, Newman D, Weaver V, Kolter R. Genetic approaches to study of biofilms. Methods Enzymol. 1999. p. 91–109.
6.
Merritt J, Kadouri D, Toole O, G. Growing and analyzing static biofilms. Curr Protoc Microbiol. 2005.
7.
Christensen G, Simpson W, Younger J, Baddour L, Barrett F, Melton D, et al. Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices. J Clin Microbiol. 1985. p. 996–1006.
8.
Hassan A, Usman J, Kaleem F, Omair M, Khalid A, Iqbal M. Evaluation of different detection methods of biofilm formation in the clinical isolates. Braz J Infect Dis. 2011. p. 305–11.
9.
Stepanović S, Vuković D, Hola V, Bonaventura G, Djukić S, Ćirković I, et al. Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. APMIS. 2007. p. 891–9.
10.
Panda P, Chaudhary U, Dube S. Comparison of four different methods for detection of biofilm formation by uropathogens. Indian J Pathol Microbiol. 2016. p. 177.
11.
Møretrø T, Hermansen L, Holck A, Sidhu M, Rudi K, Langsrud S. Biofilm formation and the presence of the intercellular adhesion locus ica among staphylococci from food and food processing environments. Appl Environ Microbiol. 2003. p. 5648–55.
12.
Mulder J, Degener J. Slime-producing properties of coagulase-negative staphylococci isolated from blood cultures. Clin Microbiol Infect. 1998. p. 689–94.
13.
Akpolat N, Elci S, Atmaca S, Akbayin H, Gül K. The effects of magnesium, calcium and EDTA on slime production byStaphylococcus epidermidis strains. Folia Microbiol. 2003. p. 649.
14.
Vasudevan P, Nair M, Annamalai T, Venkitanarayanan K. Phenotypic and genotypic characterization of bovine mastitis isolates of Staphylococcus aureus for biofilm formation. Vet Microbiol. 2003. p. 179–85.
15.
Mack D, Siemssen N, Laufs R. Parallel induction by glucose of adherence and a polysaccharide antigen specific for plastic-adherent Staphylococcus epidermidis: evidence for functional relation to intercellular adhesion. Infect Immun. 1992. p. 2048–57.
16.
Dobinsky S, Kiel K, Rohde H, Bartscht K, Knobloch J, Horstkotte M, et al. Glucose-related dissociation between ica ADBC transcription and biofilm expression by Staphylococcus epidermidis: evidence for an additional factor required for polysaccharide intercellular adhesin synthesis. J Bacteriol. 2003. p. 2879–86.
17.
Arciola C, Campoccia D, Baldassarri L, Donati M, Pirini V, Gamberini S, et al. Detection of biofilm formation in Staphylococcus epidermidis from implant infections. Comparison of a PCR-method that recognizes the presence of ica genes with two classic phenotypic methods. J Biomed Mater Res. 2006. p. 425–30.
18.
Gelosia A, Baldassarri L, Deighton M, Van Nguyen T. Phenotypic and genotypic markers of Staphylococcus epidermidis virulence. Clin Microbiol Infect. 2001. p. 193–9.
19.
Cho S, Naber K, Hacker J, Ziebuhr W. Detection of the icaADBC gene cluster and biofilm formation in Staphylococcus epidermidis isolates from catheter-related urinary tract infections. Int J Antimicrob Agents. 2002. p. 570–5.
20.
Fitzpatrick F, Humphreys H, Smyth E, Kennedy C, Gara O, J. Environmental regulation of biofilm formation in intensive care unit isolates of Staphylococcus epidermidis. J Hosp Infect. 2002. p. 212–8.
21.
Jr F, Fey V, Reller P, Chamis L, Corey A, Rupp G, et al. The intercellular adhesin locus ica is present in clinical isolates of Staphylococcus aureus from bacteremic patients with infected and uninfected prosthetic joints. Med Microbiol Immunol. 2001. p. 127–31.
22.
Frey A, Canzio D, Zurakowski J, D. A statistically defined endpoint titer determination method for immunoassays. J Immunol Methods. 1998. p. 35–41.
23.
Catic A, Gurbeta L, Kurtovic-Kozaric A, Mehmedbasic S, Badnjevic A. and Klinefelter Syndrome based on first trimester maternal serum screening data, ultrasonographic findings and patient demographics. BMC Med Genomics. 2018. p. 19.
24.
Badnjevic A, Gurbeta L, Custovic E. An expert diagnostic system to automatically identify asthma and chronic obstructive pulmonary disease in clinical settings. Sci Rep. 2018. p. 11645.
25.
Aljović A, Badnjević A, Gurbeta L. IEEE 5th Mediterranean Conference on Embedded Computing (MECO). p. 286–9.
26.
Alić B, Sejdinović D, Gurbeta L, Badnjevic A. IEEE 5th Mediterranean Conference on Embedded Computing (MECO). p. 297–300.
27.
Halilović S, Avdihodžić H, Gurbeta L. IEEE 5th Mediterranean Conference on Embedded Computing (MECO). p. 294–6.
28.
Alić B, Gurbeta L, Badnjević A. IEEE 5th Mediterranean Conference on Embedded Computing (MECO). p. 1–4.

Citation

Authors retain copyright. This work is licensed under a Creative Commons Attribution 4.0 International License. Creative Commons License

 

Article metrics

Google scholar: See link

The statements, opinions and data contained in the journal are solely those of the individual authors and contributors and not of the publisher and the editor(s). We stay neutral with regard to jurisdictional claims in published maps and institutional affiliations.