Prevalence of virulence determinants in Staphylococcus epidermidis from ICU patients in Kampala, Uganda.

INTRODUCTION: Staphylococcus epidermidis is often considered a non-pathogenic organism but it causes nosocomial infections. To distinguish invasive strains, comparative studies of patient and community isolates may offer some clues. We investigated the distribution of virulence determinants in patient isolates from Uganda. METHODOLOGY: S. epidermidis isolates were identified with the Staph API ID 32 kit. Antimicrobial susceptibility, biofilm formation and hemolysis were detected with standard procedures. Genes associated with virulence (aap, atlE, bhp, hla, hld, ica, IS256, sdrE, sea, tsst) and antimicrobial resistance (aac(6')-Ie-aph(2'')-Ia, aph(3')-IIIa, ant(4')-Ia, blaZ, mecA, vanA/vanB1) were detected by PCR. RESULTS: S. epidermidis grew in 30 (30/50, 60%) ICU samples and 20 (20/60, 33%) community samples (one isolate per sample per patient/person). All ICU isolates (30/30, 100%) were IS256 and hld positive, 22 (22/30, 73%) were biofilm/ica positive, 21 (21/30, 70%) were hemolytic on blood agar, nine (9/30, 30%) contained atlE gene, six (6/30, 20%) hla gene, five (5/30, 17%) aap gene, and three (3/30, 10%) bhp gene. A gene encoding an aminoglycoside-modifying enzyme, aph(3')-IIIa, was highly prevalent (28/30, 93%), while blaZ (2/30, 7%), mecA (3/30, 10%), vanA (3/30, 10%) and vanB1 (3/30, 10%) were less prevalent. Of the community isolates, one (1/20, 5%) was ica positive, two (2/20, 10%) formed biofilms, and three (3/20, 15%) possessed the atlE gene. bhp, aap, IS256, hld and antimicrobial resistance genes were not detected in community isolates. CONCLUSIONS: S. epidermidis from ICU patients in Mulago Hospital is potentially virulent and could be a reservoir for antimicrobial resistant genes.

Prevalence of Virulence Determinants in Staphylococcus Epidermidis from ICU Patients in Kampala; Uganda

Introduction: Staphylococcus epidermidis is often considered a non-pathogenic organism but it causes nosocomial infections. To distinguish invasive strains; comparative studies of patient and community isolates may offer some clues. We investigated the distribution of virulence determinants in patient isolates from Uganda. Methodology: S. epidermidis isolates were identified with the Staph API ID 32 kit. Antimicrobial susceptibility; biofilm formation and hemolysis were detected with standard procedures. Genes associated with virulence (aap; atlE; bhp; hla; hld; ica; IS256; sdrE; sea; tsst) and antimicrobial resistance (aac(6')-Ie-aph(2'')-Ia; aph(3')-IIIa; ant(4')-Ia; blaZ; mecA; vanA/vanB1) were detected by PCR. Results: S. epidermidis grew in 30 (30/50; 60) ICU samples and 20 (20/60; 33) community samples (one isolate per sample per patient/person). All ICU isolates (30/30; 100) were IS256 and hld positive; 22 (22/30; 73) were biofilm/ica positive; 21 (21/30; 70) were hemolytic on blood agar; nine (9/30; 30) contained atlE gene; six (6/30; 20) hla gene; five (5/30; 17) aap gene; and three (3/30; 10) bhp gene. A gene encoding an aminoglycoside-modifying enzyme; aph(3')-IIIa; was highly prevalent (28/30; 93); while blaZ (2/30; 7); mecA (3/30; 10); vanA (3/30; 10) and vanB1 (3/30; 10) were less prevalent. Of the community isolates; one (1/20; 5) was ica positive; two (2/20; 10) formed biofilms; and three (3/20; 15) possessed the atlE gene. bhp; aap; IS256; hld and antimicrobial resistance genes were not detected in community isolates. Conclusions: S. epidermidis from ICU patients in Mulago Hospital is potentially virulent and could be a reservoir for antimicrobial resistant genes

Identification of Staphylococcus aureus: DNase and Mannitol salt agar improve the efficiency of the tube coagulase test.

BACKGROUND: The ideal identification of Staphylococcus aureus clinical isolates requires a battery of tests and this is costly in resource limited settings. In many developing countries, the tube coagulase test is usually confirmatory for S. aureus and is routinely done using either human or sheep plasma. This study evaluated Mannitol salt agar and the deoxyribonuclease (DNase) test for improving the efficiency of the tube coagulase test in resource limited settings. The efficiency of human and sheep plasma with tube coagulase tests was also evaluated. METHODS: One hundred and eighty Gram positive, Catalase positive cocci occurring in pairs, short chains or clusters were subjected to growth on Mannitol salt agar, deoxyribonuclease and tube coagulase tests. Of these, isolates that were positive for at least two of the three tests (n = 60) were used to evaluate the performance of the tube coagulase test for identification of S. aureus, using PCR-amplification of the nuc gene as a gold standard. RESULTS: Human plasma was more sensitive than sheep plasma for the tube coagulase test (sensitivity of 91% vs. 81% respectively), but both plasmas had very low specificity (11% and 7% respectively). The sensitivity and specificity of the tube coagulase test (human plasma) was markedly improved when Mannitol salt agar and DNase were introduced as a tri-combination test for routine identification of Staphylococcus aureus (100% specificity and 75% sensitivity). The specificity and sensitivity of Mannitol salt agar/DNase/tube coagulase (sheep plasma) combination was 100% and 67%, respectively. CONCLUSION: The efficiency of the tube coagulase test can be markedly improved by sequel testing of the isolates with Mannitol salt agar, DNase and Tube coagulase. There is no single phenotypic test (including tube coagulase) that can guarantee reliable results in the identification of Staphylococcus aureus.