Prasanth P., Saravanakumari P.


Objective:The complications related to implant associated infections in post-angioplasty patients were considered to increase due to biofilm formation.

Methods: Genes responsible for the biofilm formation in the target organisms was investigated in the present study. The presence of the intercellular adhesion biofilm genes (icaA, icaB and icaD) was determined by the Polymerase Chain Reaction method. As preliminary investigations, standard tests, exit-site challenge test and microtitre plate method were used to study the biofilm efficiency of five different test organisms.

Results: Exit-site challenge test, was used to identify the ability of test organisms to grow on a bio-materials used in the study. Among the five selected test organisms, Staphylococcus aureus showed the highest ability to colonize the stent materials with in 24h to 48h. In microtitre plate method, Staphylococcus aureus and Staphylococcus epidermidis showed high biofilm forming index values of 0.29 and 0.27 respectively. Biofilm gene studies using PCR revealed the presence of all the three ica genes (Ica A, Ica D and, Ica B) in Staphylococcus aureus. The present research finding has great significance in the treatment to implant associated infections.

Conclusions: The results suggest that the virulence factors contributing to the development of infections can be revealed by understanding the presence of biofilm expression genes in the target organisms. This would also prevent dissemination of virulent bacteria in the health care centre; method also considered significant to detect healthy carriers of slime-producing staphylococci.


Biofilm, Exit-site, Staphylococcus aureus, icaR locus, Polymerase chain reaction

| PDF | HTML |


Wilking JN, Zaburdaev V, de Volder M, Losick R, Brenner MP, Weitz DA. Liquid transport facilitated by channels in Bacillus subtilis biofilms. Proc Natl Acad Sci 2013;110:848–52.

Thambidurai Punitha, Kannaiyan Moorthy, Ponnusamy Vijayalakshmi, Raja Vinodhini, Selvam Saranya, Murugesan Bhuvaneshwari, et al. In vitro antibacterial activity of essential plant oils against biofilm forming methicillin resistant Staphylococcus aureus. Asian J Pharm Clin Res 2014;7 Suppl 1:220-5.

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 Agric 2002;19:570-5.

Mack D. Molecular mechanisms of Staphylococcus epidermidis biofilm formation. J Hosp Infect 1999;43:113-25.

Cafiso V, Bertuccio T, Santagati M, Campanile F, Amicosante G, Perilli MG, et al. Presence of the ica operon in clinical isolates of Staphylococcus epidermidis and its role in biofilm production. Clin Microbiol Infect 2004;10:1081-8.

Boucher HW, Talbot GH, Bradley JS, Edwards JE, Gilbert D, Rice LB, et al. Bad bugs, no drugs: No eskape! An update from the infectious diseases society of America. Clin Infect Dis 2009:48:1–12.

Nasr RA, Abu Shady HM, Hussein HS. Biofilm formation and presence of icaAD gene in clinical isolates of staphylococci. Egypt J Med Human Genet 2012;13:269-74.

El Farran C, Sekar A, Balakrishnan A, Shanmugam S, Arumugam P, Gopalswamy J. Prevalence of biofilm-producing Staphylococcus epidermidis in the healthy skin of individuals in Tamil Nadu, India. Indian J Med Microbiol 2013;31:19-23.

Atshan SS, Shamsudin MN, Sekawi Z, Lung LTT, Hamat RA, Karunanidhi A, et al. Prevalence of adhesion and regulation of biofilm-related genes in different clones of Staphylococcus aureus. J Biomed Biotech 2012;5:10-7.

Agarwal A, Jain A. Glucose and sodium chloride induced biofilm production and ica operon in clinical isolates of staphylococci. Indian J Med Res 2013;138 Suppl 2:262-6.

Mertens A, Ghebremedhin B. Genetic determinants and biofilm formation of clinical Staphylococcus epidermidis isolates from blood cultures and indwelling devises. Eur J Med I 2013;3:111-9.

Bayston R, Fisher LE, Weber K. An antimicrobial modified silicone peritoneal catheter with activity against both gram positive and gram negative bacteria. Biomat 2009;30 Suppl 18:3167-73.

Christensen GD, Simpson WA, Younger JA, Baddour LM, Barrett FF, Melton DM. Adherence of coagulase negative staphylococi to plastic tissue cultures: a quantitative model for the adherence of staphylococci to medical devices. J Clin Microbiol 1985;22:996–1006.

Aricola CR, Commpaoccia D, Baldassari L, Donati ME, Gamberini S, Montanaro L. Detection of biofilm formation in Staphylococcus epidermidis from implant infections comparisons of a PCR method that recognize the presence of ica genes with two classic phenotypic methods. J Biomed Mater Res 2005;76 Suppl 2:425-30.

Angela Revelas. Healthcare–associated infections: a public health problem. Niger Med J 2012;53:59–64.

Sivarama Krishnan Rajaraman, Mallika Jainu, Gnanaprakash Dhakshinamoorthy. Ocimum Basilicum L. essential oil coated biomaterial surfaces prevent bacterial adhesion and biofilm growth. Asian J Pharm Clin Res 2016:9:379-84.

Khan F, ShuklaI RM, Mansoor T, Sharma SC. Detection of biofilm formation in Staphylococcus aureus. Does it have a role in treatment of MRSA infection? Trends Med Res 2011;10:1-8.

Seif El-Din S, El-Rehewy MS, Ghazaly MM, Abd-Elhamid MH. Biofilm formation by blood stream staphylococcal isolates from febrile pediatric cancer patients at South Egypt cancer institute. J Am Sci 2011;7 Suppl 1:674-86.

Gad GFM, El-Feky MA, El-Rehewy MS, Hassan MA, Abolella H, El-Baky HMA. Detection of icaA, icaD genes and biofilm production by Staphylococcus aureus and Staphylococcus epidermidis isolated from urinary tract catheterized patients. J Infect Dev Ctries 2009;3:342-51.

Namvar AE, Asghari B, Lari AR. Detection of the intercellular adhesion gene cluster (ica) in clinical Staphylococcus aureus isolates. Hyg Inf Cont 2013;8 Suppl 1:2196-226.

Conlon KM, Humphreys HJ. O‟Gara P. icaR encodes a transcriptional repressor involved in environmental regulation of ica operon expression and biofilm formation in Staphylococcus epidermidis. J Bacteriol 2002;184 Suppl 16:4400-8.

Murugan K, Selvanayaki K, Al-Sohaibani S. Antibiofilm activity of Andrographis paniculata against cystic fibrosis clinical isolate Pseudomonas aeruginosa. World J Microbiol Biotechnol 2011;27:1661-8.

Johannes KM, Knobloch MA, Horstkotte H, Dietrich M. Evaluation of different detection methods of biofilm formation in Staphylococcus aureus. Med Microbiol Immunol 2002;191:101-6.

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 Suppl 1:25-9.

Dobinski S, Bartscht K, Mack D. Influence of Tn917 insertion of transcription of the icaADBC operon in six biofilm-negative transposon mutants of Staphylococcus epidermidis. Plasmid 2002;47:10-7.

Oliveira A, de Lourdes RS, Cunha M. Comparison of methods for the detection of biofilm production in coagulase-negative staphylococci. BMC Res Notes 2010;1:253-60.

Gad G, El-Feky M, El-Rehewy M, Hassan M, Abolella H, Abd El-Baky R. Detection of icaA, icaD genes and biofilm production by Staphylococcus aureus and Staphylococcus epidermidisisolated from urinary tract catheterized patients. J Infect Dev Countries 2009;3 Suppl 5:342-51.

De Silva GD, Kanatazanou M, Massey RC, Wikinson AR, Day NP, Peacock SJ. The ica operon and biofilm production in coagulase negative staphylococci associated with carriage and disease in neonatal intensive care unit. J Clin Microbiol 2002;40:382-8.

Arciola CR, 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;39:2151-6.

About this article




Biofilm, Exit-site, Staphylococcus aureus, icaR locus, Polymerase chain reaction





Additional Links

Manuscript Submission


International Journal of Pharmacy and Pharmaceutical Sciences
Vol 9, Issue 11, 2017 Page: 76-80

Online ISSN


Authors & Affiliations

Prasanth P.
Research and Development Centre, Bharathiar University, Coimbatore, Tamil Nadu, India

Saravanakumari P.
Department of Microbiology, Sree Narayana Guru College, Coimbatore, Tamil Nadu, India

Article Tools


  • There are currently no refbacks.