Introduction
Materials and methods
Study area, design, and population
Collection of wound pus samples
Isolation and identification of wound bacterial isolates
Antibiotic sensitivity testing
DNA extraction and detection of virulence genes
Virulence genes | Primer Sequence | References |
---|---|---|
Hemolysin A (hla) | F: CTG ATT ACT ATC CAA GAA ATT CGA TTG R: CTT TCC AGC CTA CTT TTT TAT CAG T | [48] |
Staphylococcal enterotoxin A (sea) | F: TTG GAA ACG GTT AAA ACG AA R: GAA CCT TCC CAT CAA AAA CA | [49] |
Intracellular adhesion-A (icaA) | F: GAT TAT GTA A TG TGC TTG GA R:ACT ACT GCT GCG TTA ATA AT | [50] |
Fibronectin binding protein-A (fnbA) | F: GCG GAG ATC AAA GAC AA R: CCA TCT ATA GCT GTG TGG | [51] |
Gene | Initial Denaturation | Denaturation | Annealing | Extension | Final Extension | Cycles | Product size (bp) |
---|---|---|---|---|---|---|---|
hla | 95 °C for 5 min | 95 °C for 50 s | 58 °C for 30 s | 72 °C for 1 min | 72 °C for 10 min | 40 | 209 bp |
sea | 95 °C for 5 min | 95 °C for 1 min | 55 °C for 45 s | 72 °C for 1 min | 72 °C for 10 min | 40 | 120 bp |
icaA | 95 °C for 5 min | 95 °C for 1 min | 50 °C for 1 min | 72 °C for 1.5 min | 72 °C for 5 min | 40 | 770 bp |
fnbA | 95 °C for 5 min | 95 °C for 1 min | 47 °C for 1 min | 72 °C for 1.5 min | 72 °C for 5 min | 40 | 1279 bp |
Statistical analysis
Results
Prevalence of S. aureus isolates according to gender, age, and sample source
Antimicrobial sensitivity testing
Antibiotic | Concentration μg /disc | Sensitive No. (%) | Intermediate No. (%) | Resistant No. (%) | Company |
---|---|---|---|---|---|
Linezolid | 30 μg | 49 (83.1%) | 0 (0%) | 10 (16.9%) | Bioanalyse limited -Turkey |
Tetracycline | 30 μg | 16 (27.1%) | 6 (10.2%) | 37 (62.7%) | Himedia, India |
Chloramphenicol | 30 μg | 42 (71.2%) | 11 (18.6%) | 6 (10.2%) | Bioanalyse limited -Turkey |
Rifampin | 5 μg | 34 (57.6%) | 3 (5.1%) | 22 (37.3%) | Himedia, India |
Piperacillin | 100 μg | 0 (0%) | 0 (0%) | 59 (100%) | Bioanalyse limited -Turkey |
Gentamycin | 10 μg | 29 (49.2%) | 10 (16.9%) | 20 (33.9%) | Himedia, India |
Ampicillin / Sulbactam | 20 μg (10/10 μg) | 5 (8.5%) | 0 (0%) | 54 (91.5%) | Bioanalyse limited -Turkey |
Oxacillin | 1 μg | 5 (8.5%) | 0 (0%) | 54 (91.5%) | Sigma, USA |
Levofloxacin | 5 μg | 33(55.9%) | 5 (8.5%) | 21 (35.6%) | Himedia, India |
Ciprofloxacin | 5 μg | 31 (52.2%) | 7 (11.9%) | 21 (35.6%) | Bioanalyse limited -Turkey |
Amoxicillin/ Clavulanic | 30 μg (20/10 μg) | 5 (8.5%) | 0 (0%) | 54 (91.5%) | Bioanalyse limited -Turkey |
Vancomycin | 30 μg | 51 (86.4%) | 0 (0%) | 8 (13.5%) | Sigma, USA |
Antibiotic | Resistance profile | p* | ||
---|---|---|---|---|
Sensitive | Intermediate | Resistant | ||
Linezolid | ||||
Male | 5 | 31 | 0 | 0.111 |
Female | 5 | 10 | 0 | |
Tetracycline | ||||
Male | 25 | 7 | 4 | 0.888 |
Female | 11 | 3 | 1 | |
Chloramphenicol | ||||
Male | 6 | 25 | 5 | 0.626 |
Female | 1 | 12 | 2 | |
Rifampin | ||||
Male | 14 | 19 | 2 | 0.995 |
Female | 6 | 8 | 1 | |
Piperacillin | ||||
Male | 36 | 0 | 0 | N/A |
Female | 15 | 0 | 0 | |
Gentamycin | ||||
Male | 15 | 17 | 4 | 0.321 |
Female | 4 | 7 | 4 | |
Amoxicillin/Sulbactam | ||||
Male | 34 | 2 | 0 | 0.878 |
Female | 14 | 1 | 0 | |
Oxacillin | ||||
Male | 34 | 2 | 0 | 0.878 |
Female | 14 | 1 | 0 | |
Levofloxacin | ||||
Male | 16 | 19 | 1 | 0.579 |
Female | 5 | 10 | 0 | |
Ciprofloxacin | ||||
Male | 15 | 18 | 3 | 0.780 |
Female | 15 | 8 | 2 | |
Amoxicillin/Clavulanic acid | ||||
Male | 34 | 2 | 0 | 0.878 |
Female | 14 | 1 | 0 | |
Vancomycin | ||||
Male | 5 | 31 | 0 | 0.585 |
Female | 3 | 12 | 0 |
Antibiotic | Resistance profile | p* | ||
---|---|---|---|---|
Sensitive | Intermediate | Resistant | ||
Linezolid | ||||
1 to 20 | 4 | 19 | 0 | 0.595 |
21 to 40 | 4 | 12 | 0 | |
41 to 60 | 4 | 10 | 0 | |
Tetracycline | ||||
1 to 20 | 18 | 4 | 1 | 0.617 |
21 to 40 | 8 | 4 | 2 | |
41 to 60 | 10 | 2 | 2 | |
Chloramphenicol | ||||
1 to 20 | 2 | 19 | 2 | 0.618 |
21 to 40 | 3 | 9 | 2 | |
41 to 60 | 2 | 9 | 3 | |
Rifampin | ||||
1 to 20 | 12 | 9 | 1 | 0.395 |
21 to 40 | 3 | 10 | 1 | |
41 to 60 | 5 | 8 | 1 | |
Piperacillin | ||||
1 to 20 | 23 | 0 | 0 | N/A |
21 to 40 | 14 | 0 | 0 | |
41 to 60 | 14 | 0 | 0 | |
Gentamycin | ||||
1 to 20 | 10 | 10 | 3 | 0.851 |
21 to 40 | 4 | 8 | 2 | |
41 to 60 | 5 | 6 | 3 | |
Amoxicillin/Sulbactam | ||||
1 to 20 | 22 | 1 | 0 | 0.915 |
21 to 40 | 13 | 1 | 0 | |
41 to 60 | 13 | 1 | 0 | |
Oxacillin | ||||
1 to 20 | 22 | 1 | 0 | 0.915 |
21 to 40 | 13 | 1 | 0 | |
41 to 60 | 13 | 1 | 0 | |
Levofloxacin | ||||
1 to 20 | 11 | 12 | 0 | 0.450 |
21 to 40 | 4 | 9 | 1 | |
41 to 60 | 0 | 8 | 0 | |
Ciprofloxacin | ||||
1 to 20 | 11 | 11 | 1 | 0.439 |
21 to 40 | 5 | 8 | 1 | |
41 to 60 | 4 | 7 | 3 | |
Amoxicillin/Clavulanic acid | ||||
1 to 20 | 22 | 1 | 0 | 0.915 |
21 to 40 | 13 | 1 | 0 | |
41 to 60 | 13 | 1 | 0 | |
Vancomycin | ||||
1 to 20 | 4 | 19 | 0 | 0.556 |
21 to 40 | 1 | 13 | 0 | |
41 to 60 | 3 | 11 | 0 |
Detection of virulence genes
Virulence genes | Total No. = 59 (%) | MRSA No. = 54 (%) | MSSA No. = 5 (%) | VRSA No. = 8 (%) |
---|---|---|---|---|
hla | 22 (7.3) | 20 (36.9) | 2 (39.9) | 3 (37.5) |
sea | 43 (72.9) | 39 (72.2) | 4 (79.9) | 5 (62.5) |
icaA | 29 (49.2) | 25 (46.3) | 4 (79.9) | 4 (49.9) |
fnbA | 8 (13.6) | 6 (11.1) | 2 (39.9) | 0 (0) |
hla | P* | sea | P* | fnb | P* | icaA | P* | |||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
+ | − | + | − | + | − | + | − | |||||
Gender | ||||||||||||
Male | 16 | 20 | 0.770 | 29 | 7 | 0.125 | 6 | 30 | 0.766 | 20 | 16 | 0.311 |
Female | 6 | 9 | 9 | 6 | 2 | 13 | 6 | 9 | ||||
Age group | ||||||||||||
1 to 20 | 10 | 1 | 0.002 | 20 | 3 | 0.006 | 5 | 18 | 0.489 | 16 | 7 | 0.041 |
21 to 40 | 6 | 8 | 12 | 2 | 2 | 12 | 6 | 8 | ||||
41 to 60 | 6 | 8 | 6 | 8 | 1 | 13 | 4 | 10 |
Antibiotic | hla | sea | icaA | fnbA | ||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
R | S | p* | R | S | p* | R | S | p* | R | S | p* | |
Linzolid | ||||||||||||
Positive gene | 4 (39.9%) | 18 (36.7%) | 0.846 | 5 (50%) | 37 (75.5%) | 0.105 | 4 (39.9%) | 25 (51%) | 0.525 | 0 (0%) | 8 (16.3%) | 0.169 |
Negative gene | 6 (59.9%) | 31 (63.3%) | 5 (50%) | 12 (24.5%) | 6 (59.9%) | 24 (48.9%) | 10 (100%) | 41 (83.7%) | ||||
Tetracycline | ||||||||||||
Positive gene | 14 (37.8%) | 5 (31.3%) | 0.646 | 28 (75.7%) | 11 (68.8%) | 0.6 | 21 (56.8%) | 6 (37.5%) | 0.198 | 4 (10.8%) | 2 (12.5) | 0.602 |
Negative gene | 23 (62.2%) | 11 (68.8%) | 9 (24.3%) | 5 (31.3%) | 16 (43.2%) | 10 (62.5%) | 33 (89.2%) | 14 (87.5%) | ||||
Chloramphenicol | ||||||||||||
Positive gene | 2 (33.3%) | 14 (33.3%) | 1 | 4 (66.7%) | 30 (71.4%) | 0.81 | 3 (49.9%) | 20 (47.6%) | 0.913 | 0 (0%) | 5 (12%) | 0.372 |
Negative gene | 4 (66.7%) | 28 (66.7%) | 2 (33.3%) | 12 (28.6%) | 3 (49.9%) | 22 (52.4%) | 6 (100%) | 37 (88.1%) | ||||
Rifampin | ||||||||||||
Positive gene | 6 (27.3%) | 15 (44.1%) | 0.203 | 16 (72.7%) | 25 (73.5%) | 0.947 | 11 (49.9%) | 17 (49.9%) | 1 | 3 (13.6%) | 5 (14.7%) | 0.911 |
Negative gene | 16 (72.7%) | 19 (55.9%) | 6 (27.3%) | 9 (26.5%) | 11 (49.9%) | 17 (49.9%) | 19 (86.4%) | 29 (85.3%) | ||||
Piperacillin | ||||||||||||
Positive gene | 22 (37.3%) | 0 (0%) | < 0.0001 | 43 (72.9%) | 0 (0%) | < 0.0001 | 29 (49.2%) | 0 (0%) | < 0.0001 | 8 (13.6%) | 0 (0%) | < 0.0001 |
Negative gene | 37 (62.7%) | 0 (0%) | 16 (27.1%) | 0 (0%) | 30 (50.4%) | 0 (0%) | 51 (86.4%) | 0 (0%) | ||||
Gentamycin | ||||||||||||
Positive gene | 3 (14.9%) | 15 (51.7%) | 0.005 | 18 (89.9%) | 18 (62.1%) | 0.03 | 12 (59.9%) | 13 (44.8%) | 0.296 | 3 (14.9%) | 5 (17.2%) | 0.835 |
Negative gene | 17 (84.9%) | 14 (48.3%) | 2 (9.9%) | 11 (37.9%) | 8 (39.9%) | 16 (55.2%) | 17 (84.9%) | 17 (58.6%) | ||||
Ampicillin / Sulbactam | ||||||||||||
Positive gene | 20 (36.9%) | 2 (39.9%) | 0.896 | 39 (72.2%) | 4 (79.9%) | 0.708 | 25 (46.3%) | 4 (79.9%) | 0.044 | 6 (11.1%) | 2 (39.9%) | 0.071 |
Negative gene | 34 (62.9%) | 3 (59.9%) | 15 (27.8%) | 1 (19.9%) | 19 (35.2%) | 1 (19.9%) | 48 (88.9%) | 3 (59.9%) | ||||
Oxacillin | ||||||||||||
Positive gene | 20 (36.9%) | 2 (39.9%) | 0.896 | 39 (72.2%) | 4 (79.9%) | 0.708 | 25 (46.3%) | 4 (79.9%) | 0.044 | 6 (11.1%) | 2 (39.9%) | 0.071 |
Negative gene | 34 (62.9%) | 3 (59.9%) | 15 (27.8%) | 1 (19.9%) | 19 (35.2%) | 1 (19.9%) | 48 (88.9%) | 3 (59.9%) | ||||
Levofloxacin | ||||||||||||
Positive gene | 4 (18.9%) | 17 (51.5%) | 0.017 | 19 (90.5%) | 21 (63.6%) | 0.028 | 10 (4.8%) | 16 (48.5%) | 0.951 | 2 (9.5%) | 5 (15.2%) | 0.548 |
Negative gene | 17 (80.9%) | 16 (48.5%) | 2 (9.5%) | 12 (36.4%) | 11 (52.3%) | 17 (51.5%) | 19 (90.5%) | 28 (84.9%) | ||||
Ciprofloxacin | ||||||||||||
Positive gene | 5 (23.8%) | 13 (41.9%) | 0.178 | 20 (95.2%) | 20 (64.5%) | 0.01 | 11 (52.4%) | 14 (45.2%) | 0.609 | 4 (18.9%) | 4 (12.9%) | < 0.0001 |
Negative gene | 16 (76.2%) | 18 (58.1%) | 1 (4.8%) | 11 (35.5%) | 10 (47.6%) | 17 (54.8%) | 17 (80.9%) | 27 (87.1%) | ||||
Amoxicillin/ Clavulanic | ||||||||||||
Positive gene | 20 (36.9%) | 2 (39.9%) | 0.896 | 39 (72.2%) | 4 (79.9%) | 0.708 | 25 (46.3%) | 4 (79.9%) | 0.044 | 6 (11.1%) | 2 (39.9%) | 0.071 |
Negative gene | 34 (62.9%) | 3 (59.9%) | 15 (27.8%) | 1 (19.9%) | 19 (35.2%) | 1 (19.9%) | 48 (88.9%) | 3 (59.9%) | ||||
Vancomycin | ||||||||||||
Positive gene | 3 (37.5%) | 19 (37.3%) | 0.989 | 5 (62.5%) | 37 (72.5%) | 0.56 | 4 (49.9%) | 25 (48.9%) | 0.959 | 0 (0%) | 8 (15.7%) | 0.228 |
Negative gene | 5 (62.5%) | 32 (62.7%) | 3 (37.5%) | 14 (27.5%) | 4 (49.9%) | 26 (50.9%) | 8 (100%) | 43 84.3%) |
Virulence genes | Distribution No. (%) |
---|---|
hla + sea | 13 (21.9%) |
hla + sea + fnbA | 4 (6.8%) |
sea + fnbA | 8 (13.6%) |
hla + fnbA | 4 (6.8%) |
icaA + hla | 11 (18.6%) |
icaA + sea | 24 (40.7%) |
icaA + fnbA | 5 (8.5%) |
hla + sea + icaA + fnbA | 3 (5.1%) |