Comparative Evaluation of Methods Used For Detection of Biofilm Production In Multidrug Resistant Bacteria
Comparative Evaluation of Methods Used For Detection of Biofilm Production
DOI:
https://doi.org/10.70284/njirm.v8i5.1295Keywords:
Biofilm, Multidrug resistant bacteria, Congo red agarAbstract
Background: Multidrug resistant organisms (MDROs) are defined as bacteria resistant to at least one agent in three or more antimicrobial classes. Infections due to these bacteria lead to prolonged antimicrobial therapy and increase morbidity and mortality in the patients admitted in G.B.Pant Hospital, New Delhi. These bacteria tend to attach to various medical devices and form biofilms. Biofilm production of MDR strains has not been evaluated so far in this set up. Objective: The study was conducted to detect and compare biofilm production of MDR bacteria isolated by three different methods of biofilm detection, Congo Red Agar (CRA) method, Tube Method (TM) and Tissue Culture Method (TCP). Methods: A total of 200 bacterial isolates (MDROs) from various clinical samples of patients suffering from hospital acquired infections were subjected to biofilm detection along with positive and negative controls by all three methods (TCP, TM, and CRA). Result: Of the 200 isolates (MDROs), 152 (76%) were biofilm producers i.e. strong 28 (14%), moderate 57(28.5%), weak 67 (33.5%) by TCP Method. By TM 166 (83%) were biofilm producers i.e strong 34 (17%), moderate 86 (43%) and weak 46 (23%). By CRA Method 126 (63%) isolates were biofilm producers and 74 were non-biofilm producers. False positive rate was 15% and 16.5% and false negative rate was 8% and 29.5% respectively by TM and CRA methods, taking in to consideration TCP as a Gold Standard method of biofilm detection.11 Conclusion: Tube Method has detected the highest number of biofilm producers followed by TCP and CRA. But Tube Method (TM) cannot be considered as most reliable method as the interpretation of the result depends upon the individual’s observations. False negativity was high in case of CRA method. So it is concluded that TCP is the most reliable quantitative method of screening of biofilm production of bacterial isolates in Laboratories. [Gyaneshwar T NJIRM 2017; 8(5):1-8]
References
2. Magiorakos A.P, Srinivasan A, Carey R.B, Carmeli Y, Falagas M.E, Giske C.G. et al. Multidrug resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance, Clinical Microbiol Infection 2012;18: 268-281.
3. Bergogne-Berezin E, Decre D, Joly-Guillou ML. Opportunistic nosocomial multiply resistant bacterial infections- their treatment and prevention. J Antimicrob Chemother. 1993;32(Suppl A): 39-47.
4. McGrath EJ, Asmar BI. Nosocomial infections and multidrug-resistant bacterial organisms in the pediatric intensive care unit. Indian J Pediatr. 2011;78: 176-184.
5. Revdiwala S, Rajdev BM, Mulla S. Characterization of bacterial etiologic agents of biofilm formation in medical devices in critical care set up. Crit. Care Res. Pract. 2012; 945-805.
6. Arciola C. R, Baldassarri L, Montanaro L. Presence of icaA and icaD genes and slime production in a collection of Staphylococcal strains from catheter associated infections. J. Clin. Microbiol.2001; 39: 2151–2156.
7. Research on microbial biofilms (PA-03-047). NIH, National Heart, Lung and Blood Institute. 2002; p12-20.
8. Donlan RM. Biofilms and device-associated infections. Emerg. Infect. Dis. 2001; 7(2): 277-818.
9. Freeman J, Falkiner FR, Keane CT. New method for detecting slime production by coagulase negative Staphylococci. J. Clin. Pathol.1989; 42:872-4.
10. Christensen GD, Simpson WA, Younger JJ et al. Adherence of coagulase negative Staphylococci to plastic tissue cultures: a quantitative model for the adherence of Staphylococci to medical devices. J. Clin. Microbiol.1985; 22:996-1006.
11. Mathur T, Singhal S, Khan S, Upadhyay DJ, Fatma T, Rattan A. Detection of biofilm formation among the clinical isolates of Staphylococci: an evaluation of three different screening methods. Indian J. Med. Microbiol.2006; 24(1):25-9.
12. Donlan RM, Murga R, Bell M et al. Protocol for detection of biofilms on needleless connectors attached to central venous catheters. J Clin Microbiol.2001; 39:750-3.
13. Aparna MS, Yadav S. Biofilms: microbes and disease. Braz. J Infect Dis. 2008; 12(6):526-30.
14. Zufferey J, Rime B, Francioli P, Bille J. Simple method for rapid diagnosis of catheter associated infection by direct Acridine orange staining of catheter tips. J Clin Microbiol.1988; 26:175-7.
15. Bauer AW, Kirby M M, Sherris JC, Jurek M. Antibiotic susceptibility testing by a standardized single method. Am J Clin Pathol.1966; 45:493-6.
16. Stepanovic S, Vukovi D, Hola V et al. Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by Staphylococci. APMIS. 2007; 115:891-9.
17. Reid G. Biofilms in infectious disease and on medical devices. Int. J. Antimic. Ag. 1999; 11:223-6.
18. Kim L. Riddle of biofilm resistance. Antimic Ag chemother. 2001; 45(4):999-1007.
19. Fatima S, Prasanthi K, Nagamani K. Comparative evaluation of biofilm production in Multidrug resistant and sensitive Gram negative clinical isolates Int.J.Curr.Microbiol.App.Sci. 2015; 4(6): 918-926.
20. Gurung J, Khyriem A.B, Banik A, et al. Association of biofilm production with multidrug resistance among clinical isolates of Acinetobacter baumannii and Pseudomonas aeruginosa from intensive care unit. Indian J Crit Care Med. 2013 Jul-Aug; 17(4): 214–218.
21. Dijkshoorn L, Nemec A, Seifert H. An increasing threat in hospitals: Multidrug resistant Acinetobacter baumannii. Nat Rev Microbiol. 2007; 5: 939-51.
22. Navon-Venezia S, Ben-Ami R, Carmeli Y. Update on Pseudomonas aeruginosa and Acinetobacter baumannii infections in the health care setting. Curr Opin Infect Dis. 2005; 18:306-13.
23. Hassan A, Usman J, Kaleem F, et al. Evaluation of different detection methods of biofilm formation in the clinical isolates. Braz J Infect Dis. 2011;15(4):305-311.
24. Nagaveni S, Rajeshwari H, Oli A. K, et al. Evaluation of biofilm forming ability of the multi-drug resistant Pseudomonas aeruginosa. The Bioscan, 2010;5(4):563-566.
25. Rao RS, Karthika RU, Singh SP, Shashikala P, Kanungo R, Jaychandran S, et al, Correlationbetween biofilm production and multi drug resistance in Imipenem resistant clinical isolates of Acinetobacter baumanii. Indian J Med Microbiol.2008; 26:333-7.
26. Rodriguez-Bano J, Marti S, Soto S, Fernandez-Cuenca F, Cisneros JM, Pachon J, et al. Spanish group for the study of Nosocomial Infections (GEIH). Biofilm formation in Acinetobacter baumanii, associated features and clinical implications. Clin Microbiol Infect. 2008; 14:276-8.
27. Marti S, Rodriguez-Bano J, Catel-Ferreira M, Jouenne T, Vila J, Seifert H, et al. Biofilm formation at the solid- liquid and air-liquid interfaces by Acinetobacter species. BMC Res Notes. 2011; 4:5.
28. Ruzika F, Hola V, Votava M, et al.Biofilm detection and clinical significance of Staphylococcus epidermidis isolates. Folia Microbiol (Prabha).2004;49(5):596-600. 29. Baqai R, Aziz M, Rasool G. Urinary tract infection in diabetic patients and biofilm formation of uropathogens. Infect Dis J Pakistan. 2008; 17(1):7-9.
30. Christensen GD, Simpson WA, Bisno AL, Bachey EH, Adherence of slime producing strains of Staphylococcus epidermidis to smooth surfaces. Infect Immun 1982; 37:318-26.
31. Zaranza A.V, Morais F.C, Carmo M.S, et al. Antimicrobial Susceptibility, Biofilm production and Adhesion to HEp-Cells of Pseudomonas aerigunosa strains isolated from Clinical Samples. Journal of Biomaterials and Nanobiotechnology, 2013; 4: 98-106.
32. Knobloch JK, Horsetkotte MA, Rohde H, Mack D. Evaluation of different detection methods of biolfilm formation in Staphylococcus aureus. Med Microbial Immunol 2002; 191(2):101-6.
