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Vol. 15. Issue 4.
Pages 305-311 (July - August 2011)
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Vol. 15. Issue 4.
Pages 305-311 (July - August 2011)
Original article
Open Access
Evaluation of different detection methods of biofilm formation in the clinical isolates
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Afreenish Hassan1,
Corresponding author
afreenish216a@yahoo.com

Correspondence to: Department of Microbiology National University of Sciences and Technology, Islamabad, Army Medical College, Rawalpindi, Pakistan.
, Javaid Usman2, Fatima Kaleem1, Maria Omair3, Ali Khalid1, Muhammad Iqbal4
1 MPhil Microbiology, Department of Microbiology, National University of Sciences and Technology (NUST), Islamabad, Army Medical College, Rawalpindi, Pakistan
2 Professor of Pathology, Department of Microbiology, NUST, Islamabad, Army Medical College, Rawalpindi, Pakistan
3 FCPS Microbiology, Department of Microbiology, NUST, Islamabad, Army Medical College, Rawalpindi, Pakistan
4 ST, Senior Technician, Department of Microbiology, NUST, Islamabad, Army Medical College, Rawalpindi, Pakistan
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Abstract
Background

Microorganisms growing in a biofilm are associated with chronic and recurrent human infections and are highly resistant to antimicrobial agents. There are various methods to detect biofilm production like Tissue Culture Plate (TCP), Tube method (TM), Congo Red Agar method (CRA), bioluminescent assay, piezoelectric sensors, and fluorescent microscopic examination.

Objective

This study was conducted to compare three methods for the detection of biofilms.

Method

The study was carried out at the Department of Microbiology, Army Medical College, National University of Sciences and Technology, Pakistan, from January 2010 to June 2010. A total of 110 clinical isolates were subjected to biofilm detection methods. Isolates were identified by standard microbiological procedures. Biofilm detection was tested by TCP, TM and CRA. Antibiotic susceptibility test of biofilm producing bacteria was performed by using the Kirby-Bauer disc diffusion technique according to CLSI guidelines.

Results

The TCP method was considered to be superior to TM and CRA. From the total of 110 clinical isolates, TCP method detected 22.7% as high, 41% moderate and 36.3% as weak or non-biofilm producers. We have observed higher antibiotic resistance in biofilm producing bacteria than non-biofilm producers.

Conclusion

We can conclude from our study that the TCP method is a more quantitative and reliable method for the detection of biofilm forming microorganisms as compared to TM and CRA methods, and it can be recommended as a general screening method for detection of biofilm producing bacteria in laboratories.

Keywords:
biofilms
bacteria
anti-bacterial agents
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References
[1.]
R.M. Donlan, W. Costerton.
Biofilms: Survival mechanisms of clinically relevant Microorganisms.
Clin Microbiol Rev, 15 (2002), pp. 167-193
[2.]
D. Thomas, F. Day.
Biofilm formation by plant associated bacteria.
Ann Rev Microbiol, 61 (2007), pp. 401-422
[3.]
P.S. Stewart, J.W. Costerton.
Antibiotic resistance of bacteria in biofilms.
Lancet, 358 (2001), pp. 135-138
[4.]
Research on microbial biofilms (PA-03-047). NIH, National Heart, Lung, and Blood Institute. 2002-12-20.
[5.]
G. Reid.
Biofilms in infectious disease and on medical devices.
Int. J. Antimic Ag, 11 (1999), pp. 223-226
[6.]
R.M. Donlan.
Biofilms and device-associated infections.
Emerg Infect Dis, 7 (2001), pp. 277-281
[7.]
G.D. Christensen, W.A. Simpson, J.A. Younger, 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, 22 (1995), pp. 996-1006
[8.]
G.D. Christensen, W.A. Simpson, A.L. Bisno, E.H. Beachey.
Adherence of slime producing strains of Staphylococcus epidermidis to smooth surfaces.
Infect Immun, 37 (1982), pp. 318-326
[9.]
J. Freeman, F.R. Falkiner, C.T. Keane.
New method for detecting slime production by coagulase negative staphylococci.
J Clin Pathol, 42 (1989), pp. 872-874
[10.]
R.M. Donlan, R. Murga, M. Bell, et al.
Protocol for detection of biofilms on needleless connectors attached to central venous catheters.
J Clin Microbiol, 39 (2001), pp. 750-753
[11.]
M.S. Aparna, S. Yadav.
Biofilms: microbes and disease.
Braz J Infect Dis, 12 (2008), pp. 526-530
[12.]
J. Zufferey, B. Rime, P. Francioli, J. Bille.
Simple method for rapid diagnosis of catheter associated infection by direct Acridine orange staining of catheter tips.
J Clin Microbiol, 26 (1988), pp. 175-177
[13.]
T. Mathur, S. Singhal, S. Khan, D.J. Upadhyay, T. Fatma, A. Rattan.
Detection of biofilm formation among the clinical isolates of staphylococci: an evaluation of three different screening methods.
Indian J Med Microbiol, 24 (2006), pp. 25-29
[14.]
S. Stepanovic, D. Vukovi, V. Hola, et al.
Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by Staphylococci.
[15.]
A.W. Bauer, M.M. Kirby, J.C. Sherris, M. Jurek.
Antibiotic susceptibility testing by a standardized single method.
Am J Clin Pathol, 45 (1966), pp. 493-496
[16.]
L. Kim.
Riddle of biofilm resistance.
Antimic Ag Chemother, 45 (2001), pp. 999-1007
[17.]
F. Eftikhar, D.P. Speert.
Biofilm formation by persistent and non-persistent isolates of Staphylococcus epidermidis from a neonatal intensive care unit.
J Hosp Infect, 71 (2009), pp. 112-116
[18.]
S. Bose, M. Khodke, S. Basak, S.K. Mallick.
Detection of biofilm producing staphylococci: need of the hour.
J Clin Diagn Res, 3 (2009), pp. 1915-1920
[19.]
F. Ruzicka, V. Hola, M. Votava, et al.
Biofilm detection and clinical significance of Staphylococcus epidermidis isolates.
Folia Microbiol (Praha), 49 (2004), pp. 596-600
[20.]
R. Baqai, M. Aziz, G. Rasool.
Urinary tract infection in diabetic patients and biofilm formation of uropathogens.
Infect Dis J Pakistan, 17 (2008), pp. 7-9
[21.]
J.K. Knobloch, M.A. Horsetkotte, H. Rohde, D. Mack.
Evaluation of different detection methods of biolfilm formation in Staphylococcus aureus.
Med Microbial Immunol, 191 (2002), pp. 101-106
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