Coagulase negative staphylococci in Anti-Cancer Center, Batna, Algeria: antibiotic resistance pattern, biofilm formation, and detection of mecA and icaAD genes

Abstract

Background: Coagulase-negative staphylococci (CoNS) are normal microbial flora found on the skin and mucous membranes of mammals. Considered for a long time as avirulent commensals, these bacteria are now recognized as opportunistic pathogens by virtue of their high resistance to multiple antibiotics and capacity for biofilm formations, which made them important agents of nosocomial and community-acquired infections. The objectives of this study are to determine the antibiotic resistance pattern and biofilm formation, and to detect mecA and icaAD genes in clinical CoNS isolates from Batna’s Anti-Cancer Center (ACC) in Algeria.

Methods: A total of 66 CoNS were isolated from different samples and identified by API Staph system. In vitro antibiotic susceptibility testing (AST) of each isolate to selected antibiotics was determined by the disk diffusion method, and minimum inhibitory concentrations (MICs) of oxacillin and vancomycin were determined by E-test. Biofilm formation was assessed by Tissue Culture Plate (TCP) and Congo Red Agar (CRA) methods. The polymerase chain reaction (PCR) was used to amplify mecA gene in 9 oxacillin-resistant and 1 oxacillinsensitive CoNS, and icaAD gene in 9 biofilm forming and 1 non-biofilm forming CoNS. Sequencing of the 16S rDNA of 1 mecA and 1 icaAD positive isolates was performed by the Sanger method.

Results: Nine species of CoNS were identified, with Staphylococcus epidermidis (n=29, 44%) and Staphylococcus haemolyticus (n=15, 22.7%) constituting the largest proportion, and isolated mainly from the onco-haematology service unit of the center. The isolates were resistant to penicillin G (98.5%), cefoxitin (80.3%) and oxacillin (72.2%). The TCP method was more sensitive (89.4%) than CRA method (31.8%) in detecting biofilm formation. The mecA gene was detected in 66.7% (6/9) of oxacillin resistant CoNS and the icaAD gene in 55.6% (5/9) of TCP positive CoNS isolates

Conclusion: Invitro resistance to methicillin (oxacillin) and biofilm formation were high among the CoNS isolates in this study, but the association of these with respective carriage of mecA and icaAD genes was low.

Keywords: Coagulase negative staphylococci, identification, antibiotic resistance, biofilm, PCR