High prevalence of toxic shock syndrome toxin-producing epidemic methicillin-resistant Staphylococcus aureus 15 (EMRSA-15) strains in Kuwait hospitals.

This study characterized EMRSA-15 isolates obtained from patients in Kuwait hospitals for their genotypic relatedness, antibiotic resistance and carriage of virulence genes using pulsed-field gel electrophoresis (PFGE), coagulase serotyping, SCCmec subtyping, spa typing, multilocus sequence typing and DNA microarray. The isolates were resistant to trimethoprim (75.6%), ciprofloxacin (29.7%), erythromycin and clindamycin (24.3%), tetracycline (19.0%), and gentamicin and kanamycin (21.6%). All 37 isolates belonged to sequence type (ST) 22, coagulase type XI, three PFGE types and eight subtypes, ten spa types including t223 (51.3%), t852 (13.5%), t032 (8.1%), t790 (8.1%), t3107 (5.4%) and one each of t309, t2251, t3935, t5708 and t5983. Twenty-six isolates (70.2%) carried SCCmec IVa, eight isolates carried SCCmec IV and three isolates carried SCCmec IVh. All isolates carried agr1, cap5 and egc gene cluster (seg, sei, selm, seln, selo, and selu). tst (toxic shock syndrome toxin) was detected in 23 isolates. Eight isolates (21.6%) were positive for Panton-Valentine leukocidin (PVL). Genotypic analysis revealed that 62.1% of the isolates comprising ST22-IVa-t223 (51.3%) and ST22-IVa-t309/t2251/t3935/t5708 (10.8%) were CC22-[tst1(+)] UK EMRSA-15/Middle Eastern variant, 21.6% were CC22-PVL(+) EMRSA-15 variant and 16.2% were CC22-UK EMRSA-15/Barnim clone. These results show that the tst1 positive-ST22-IVa-t223 (Middle Eastern variant) and the CC22-PVL(+) EMRSA-15 variant were the dominant EMRSA-15 variants in Kuwait hospitals.

Dissemination of methicillin-resistant Staphylococcus aureus SCCmec type IV and SCCmec type V epidemic clones in a tertiary hospital: challenge to infection control.

Two-hundred MRSA strains from inpatients with healthcare-associated (HA) and 100 MRSA strains from outpatients with community-associated (CA) skin and soft tissue infections (SSTIs) were tested for antimicrobial susceptibility, staphylococcal cassette chromosome mec (SCCmec) typing, Panton-Valentine leucocidin (PVL) toxin, seh and arcA genes. Based on SCCmec typing, HA-MRSA isolates were further divided into HA-SCCmec I/II/III MRSA and HA-SCCmec IV/V MRSA, and CA-MRSA isolates into CA-SCCmec I/II/III MRSA and CA-SCCmec IV/V MRSA. SCCmec types were further characterized by pulsed-field gel electrophoresis, spa typing and multi-locus sequence typing. Seventy-five (37·5%) HA-MRSA isolates and 83/100 CA-MRSA isolates were SCCmec IV/V genotype. HA-SCCmec IV/V MRSA was associated with malignancy (P = 0·03) and bone fractures (P = 0·02) compared to CA-SCCmec IV/V MRSA. HA-SCCmec IV/V MRSA was associated with PVL gene carriage compared to HA-SCCmec I/II/III MRSA (P < 0·001). ST22-MRSA-IV (EMRSA-15), ST772-MRSA-V, and ST36-MRSA-IV and ST239:EMRSA-I:III were the major clones identified. Our study documents the emergence of SCCmec IV and SCCmec V MRSA clones in an Indian hospital.

Genotyping of methicillin-resistant Staphylococcus aureus in the Sultan Qaboos University Hospital, Oman reveals the dominance of Panton-Valentine leucocidin-negative ST6-IV/t304 clone.

The objective of this study was to determine the prevalence and distribution of methicillin-resistant Staphylococcus aureus (MRSA) genotypes circulating at a tertiary hospital in the Sultanate of Oman. A total of 79 MRSA isolates were obtained from different clinical samples and investigated using antibiogram, pulsed-field gel electrophoresis (PFGE), staphylococcal chromosome cassette mec (SCCmec), Spa typing and multilocus sequence typing (MLST). The isolates were susceptible to linezolid, vancomycin, teicoplanin, tigecycline and mupirocin but were resistant to tetracycline (30.4%), erythromycin (26.6%), clindamycin (24.1%), trimethoprim (19.0%), ciprofloxacin (17.7%), fusidic acid (15.2%) and gentamicin (12.7%). Molecular typing revealed 19 PFGE patterns, 26 Spa types and 21 sequence types. SCCmec-IV (86.0%) was the dominant SCCmec type, followed by SCCmec-V (10.1%). SCCmec-III (2.5%) and SCCmec-II (1.3%) were less common. ST6-IV/t304 (n = 30) and ST1295-IV/t690 (n = 12) were the dominant genotypes followed by ST772-V/t657 (n = 5), ST30-IV/t019/t021 (n = 5), ST22-IV/t852 (n = 4), ST80-IV/t044 (n = 3) and 18 single genotypes that were isolated sporadically. On the basis of SCCmec typing and MLST, 91.2% of the isolates were classified as community-associated MRSA and 8.8% of the isolates (consisting of four ST22-IV/t852, one ST239-III/t632, one ST5-III/t311 and one ST5-II/t003) were classified as healthcare-associated MRSA. The study has revealed the dominance of a Panton-Valentine leucocidin-negative ST6-IV/t304 clone and provided insights into the distribution of antibiotic resistance in MRSA at the tertiary hospital in Oman. It also highlights the importance of surveillance in detecting the emergence of new MRSA clones in a healthcare facility.

A case of community-onset meningitis caused by hospital methicillin-resistant Staphylococcus aureus successfully treated with linezolid and rifampicin.

OBJECTIVE: To report a relatively rare presentation of methicillin-resistant Staphylococcus aureus (MRSA) meningitis in a previously healthy boy in Kuwait. CLINICAL PRESENTATION AND INTERVENTION: A 14-year-old boy presented with a 2 weeks' history of headache and fever with increasing severity. He developed photophobia and double vision 2 days prior to his hospital visit and received ceftriaxone for 6 days prior to admission to the hospital. There was no history of head trauma or neurosurgical operation. Lumbar puncture revealed a slightly turbid cerebrospinal fluid with pleocytosis and greatly reduced glucose, elevated protein level and on culture grew MRSA. Staphylococcal chromosome cassette mec (SCCmec) typing revealed that it belonged to SCCmec type III and sequence type 238 (ST238-SCCmec-III). Polymerase chain reaction screening for the presence of Panton-Valentine leukocidin (PVL) genes yielded a negative result; all these findings were consistent with hospital-acquired MRSA. He was treated with intravenous linezolid and rifampicin for 2 weeks, made good response and was discharged home fully recovered and well. CONCLUSION: Hospital MRSA should be considered in the differential diagnosis of the causative agents of community-onset meningitis in healthy patients even without predisposing factor.

Clinical and molecular characteristics of nosocomial meticillin-resistant Staphylococcus aureus skin and soft tissue isolates from three Indian hospitals.

We analysed risk factors for nosocomial meticillin-resistant Staphylococcus aureus (MRSA) skin and soft tissue infections (SSTIs) in three Indian hospitals. We also determined antimicrobial resistance patterns and genotypic characteristics of MRSA isolates using pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and staphylococcal cassette chromosome (SCCmec) typing. Medical records of 709 patients admitted to three tertiary hospitals with nosocomial S. aureus SSTIs were clinically evaluated. Antimicrobial susceptibility testing of patient isolates was performed in accordance with Clinical and Laboratory Standards Institute guidelines, with meticillin and mupirocin resistance confirmed by multiplex polymerase chain reaction. PFGE analysis of 220 MRSA isolates was performed, followed by MLST and SCCmec typing of a selected number of isolates. MRSA was associated with 41%, 31% and 7.5% of infections at the three hospitals, respectively. Multiple logistic regression analysis identified longer duration of hospitalisation [odds ratio (OR): 1.78; OR: 2.83 for >or=20 days], intra-hospital transfer (OR: 1.91), non-infectious skin conditions (3.64), osteomyelitis (2.9), neurological disorders (2.22), aminoglycoside therapy (1.74) and clindamycin therapy (4.73) as independent predictors for MRSA SSTIs. MRSA isolates from all three hospitals were multidrug resistant, with fifteen clones (I-XV) recognised. A majority of the strains possessed type III cassette. The common sequence type (ST) 239 was considered the signature MLST sequence for PFGE clone III. This major MRSA clone III was closely related to the UK EMRSA-1 and was significantly more resistant to antibiotics. Dissemination of multidrug-resistant MRSA clones warrants continuous tracking of resistant genotypes in the Indian subcontinent.

Genetic lineages of community-associated methicillin-resistant Staphylococcus aureus in Kuwait hospitals.

Twenty-six community-associated methicillin-resistant Staphylococcus aureus (CAMSRA) isolates were characterized by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) and screened for accessory gene regulator (agr), capsular polysaccharide (cap), and Panton-Valentine leucocidin (PVL) genes. They exhibited five PFGE patterns (types A to E). The majority were PFGE type A (12 isolates) or type B (8 isolates). MLST showed that PFGE type A isolates belonged to sequence type 80 (ST80), while the PFGE type B isolates were ST30. The ST80 and ST30 clones contained agr allotype 3, cap type 8, and PVL. The results showed that two internationally recognized CAMRSA clones are dominant in Kuwait hospitals.

Surveillance of antibacterial resistance in Staphylococcus aureus isolated in Kuwaiti hospitals.

OBJECTIVE: To investigate the prevalence of antibiotic resistance among Staphylococcus aureus isolated in Kuwaiti hospitals. MATERIALS AND METHODS: S. aureus were isolated and identified following standard microbiological methods. Antibacterial susceptibility test was performed by disk diffusion and the measurement of minimum inhibitory concentration with E-test strips. RESULTS: A total of 1,846 S. aureus isolates were analyzed from 13 hospitals between 1 March and 30 October 2005. They were isolated from 1,765 (95.6%) inpatients and 81 (4.4%) outpatients. Methicillin resistance was detected in 588 (32.0%) of the isolates. The methicillin-resistant S. aureus (MRSA) consisted of 461 (78%) multiresistant and 127 (22%) nonmultiresistant isolates. The nonmultiresistant MRSA consisted of epidemic MRSA-15 and community-associated MRSA. The community-associated MRSA was detected in all hospitals with MRSA, indicating its establishment in Kuwaiti hospitals. The proportion of isolates resistant to gentamicin, kanamycin, erythromycin, tetracycline, ciprofloxacin, fusidic acid and trimethoprim was higher among MRSA than methicillin-susceptible S. aureus (MSSA) isolates. Twenty-four and 22% of MRSA and MSSA isolates, respectively, expressed reduced susceptibility to vancomycin (minimum inhibitory concentration = 3-4 mg/l). CONCLUSION: The study revealed the presence of methicillin resistance in 32% of S. aureus isolated in Kuwaiti hospitals and revealed an increase in the number of MRSA and MSSA with reduced susceptibility to vancomycin.

Plasmid-mediated high-level ceftriaxone resistance in a Salmonella enterica serotype typhimurium isolate.

OBJECTIVE: To present the first documented case of acute infectious gastroenteritis caused by high-level ceftriaxone-resistant Salmonella enterica serotype typhimurium in Kuwait. SUBJECT AND METHODS: Isolation from stool specimen and species identification of current enteric pathogen was carried out according to standard methods. Susceptibility to antibiotics was determined by the disc diffusion method on Mueller-Hinton agar. Minimal inhibitory concentrations (MICs) were measured with E-test strips. The production of extended spectrum beta-lactamase (ESBL) was studied by the double disc synergy method and E-test ESBL strips. Plasmid DNA isolation was performed by the rapid alkaline lysis method. Plasmid DNA was transferred by conjugation to a recipient strain of Escherichia coli. RESULTS: The isolate of S. enterica serotype typhimurium was resistant to ceftriaxone (MIC >256 mg/l), cefotaxime and ceftazidime, and produced ESBL. Ceftriaxone and cefotaxime resistance were co-transferred on a 3.2-kb plasmid to the E. coli recipient strain. Loss of the 3.2-kb plasmid from the transconjugant resulted in the co-loss of ceftriaxone and cefotaxime resistance confirming the carriage of ceftriazone resistance on the 3.2-kb plasmid. CONCLUSION: Plasmid-mediated high-level resistance to ceftriaxone and ESBL production in Salmonella serotype typhimurium is an emerging problem among Salmonella that requires closer monitoring of antimicrobial resistance among these bacterial species.

Characterisation of non-multiresistant methicillin-resistant Staphylococcus aureus (including EMRSA-15) in Kuwait Hospitals.

This study characterised non-multiresistant methicillin-resistant Staphylococcus aureus (nmMRSA) isolates from Kuwait hospitals to ascertain whether they were community-acquired MRSA (CA-MRSA). Forty-two nmMRSA isolates obtained between July 2001 and October 2003 were analysed by staphylococcal cassette chromosome mec (SCCmec) typing, bacteriophage typing, production of Panton-Valentine leukocidin (PVL), urease and staphylococcal enterotoxins A, B, C and D, TSST-1, and by pulsed-field gel electrophoresis (PFGE). Forty-one isolates were SCCmec type IV, and one isolate was SCCmec type III. The isolates belonged to six PFGE patterns, with two types, A and D, distributed in six and four hospitals, respectively. Most (n = 26; 61.9%) isolates produced urease. These isolates were mainly from wound and skin infections, showed low-level methicillin resistance (MIC 8-48 mg/L), and nine carried genes for PVL. These characteristics, together with their carriage of the type-IV SCCmec, identified the isolates as CA-MRSA. Ten of the 16 urease-negative isolates produced staphylococal enterotoxin C; 12 reacted weakly with phage 75, and were resistant to clindamycin and/or erythromycin, which are characteristics of EMRSA-15. Thus, this study identified the co-existence of two types of nmMRSA, i.e., CA-MRSA and EMRSA-15, in Kuwait hospitals.

Antibacterial resistance and molecular typing of methicillin-resistant Staphylococcus aureus in a Kuwaiti general hospital.

OBJECTIVE: To investigate antibiotic resistance and genetic relatedness of methicillin-resistant Staphylococcus aureus (MRSA) isolated in a general hospital in Kuwait over a period from 1996 to 1998 and 2001. MATERIAL AND METHODS: The isolates were characterized by antibacterial susceptibility testing, coagulase serotyping, coagulase gene polymorphism (coag-RFLP) and pulsed-field gel electrophoresis (PFGE). RESULTS: The MRSA isolates were highly resistant to gentamicin, kanamycin, ciprofloxacin, tetracycline, fusidic acid and mupirocin. The prevalence of gentamicin, kanamycin, streptomycin, tetracycline and erythromycin resistance remained high (80-96%) throughout the study period, but the prevalence of resistance to ciprofloxacin, fusidic acid and mupirocin steadily increased. The already high mupirocin resistance level increased from 12.5 in 1996, to 85.7% in 2001, and the fusidic acid resistance varied between 70.8 and 85.7%. In contrast, chloramphenicol and trimethoprim resistance declined from 25 and 29% in 1996 to 4.7 and 14.2% in 2001, respectively. The majority (91.5%) of the isolates were coagulase serotype 4. AluI restriction endonuclease analysis of amplified coagulase gene generated four coag-RFLP patterns: 92% of them were coag-RFLP type 1, while types 2, 3 and 4 were 3.5, 4.6 and 1.1% respectively. PFGE differentiated them into seven pulsotypes (PFGE types 1-7). The PFGE type 1 pulsotype constituted 90.2% of the isolates. Isolates with the type A coag-RFLP also had the type1 PFGE pulsotypes. CONCLUSION: The concordant results of PFGE and coag-RFLP demonstrated the presence of a persistent MRSA clone in the hospital during the study period.

Etiology and antibiotic susceptibility patterns of community-acquired urinary tract infections in a Kuwait hospital.

OBJECTIVE: The aim of this study was to determine the distribution and antibiotic susceptibility patterns of bacterial strains isolated from patients with community-acquired urinary tract infections (UTIs) at the Infectious Diseases Hospital, Kuwait. MATERIALS AND METHODS: The study was conducted over a 7-year period. Patient information was obtained from medical record files. Antibiotic-sensitivity testing was performed by disk diffusion. E test and double disk diffusion methods were used to study the production of extended spectrum beta-lactamases. RESULTS: Of the 14,042 urine samples processed, significant bacteriuria (>10(5) cfu/ml) was detected in 1,606 (11.4%). The majority (74.5%) of the isolates were from women while the remaining 25.5% were from men. The majority of infections (75%) were due to Enterobacteriaceae, coagulase-negative staphylococci (10.3%) and group B streptococci (8.7%). Among the gram-negative enteric bacilli high prevalence of resistance to ampicillin, amoxicillin/clavulanic acid, cephalothin, and trimethoprim/sulfamethoxazole was observed. Increasing resistance to ciprofloxacin and gentamicin was observed in E. coli isolates over the 7 years. Multiple resistance was detected in 53.8 and 41% of E. coli and Klebsiella spp. strains, respectively. No glycopeptide-resistant enterococci were isolated. CONCLUSION: This study revealed that Enterobacteriaceae were the predominant bacterial pathogen of community-acquired UTIs in Infectious Diseases Hospital, Kuwait. It also demonstrated an increasing resistance to ciprofloxacin, gentamicin and the production of extended spectrum beta-lactamase among UTI pathogens in the community.

Characterization of high-level aminoglycoside-resistant enterococci in Kuwait hospitals.

This study investigated the distribution of genes for aminoglycoside-modifying enzymes (AME) and the genetic relatedness of high-level aminoglycoside-resistant enterococci isolated in Kuwait hospitals. A total of 117 enterococci, consisting of 109 Enterococcus faecalis, seven Enterococcus faecium, and one Enterococcus casseliflavus were studied. The MICs of gentamicin, kanamycin, amikacin, tobramycin, and streptomycin were determined by agar dilution and the genes encoding the AAC(6')- APH(2"), ANT(4'), APH(3'), APH (2")-Ib, APH (2")-Ic, APH (2")-Id, and ANT(6) enzymes were amplified by PCR. They were typed by pulsed-field gel electrophoresis (PFGE). Filter mating was used to transfer gentamicin resistance determinants. They were all resistant to kanamycin (MIC 2000 mg/L). Fifty-five isolates were resistant to gentamicin (MIC 500 mg/L), 72 were resistant to tobramycin (MIC 64 mg/L), 115 were resistant to amikacin (MIC 64 mg/L), and 97 were resistant to streptomycin (MIC 1000 mg/L). The aac(6')-Ie-aph(2")-Ia was detected in all isolates with gentamicin MIC 500 mg/L and in 15 isolates with gentamicin MIC 256 mg/L. The aph(3')-IIIa gene was detected in 101 isolates, whereas the ant(6')-Ia gene was detected in 85 of the 97 streptomycin-resistant isolates with MIC 1000 mg/L. The aac(6')-Ii gene was detected only in the seven E. faecium isolates. None of them contained ant(4')-Ia, aph(2")-Ib, aph(2")-Ic and aph(2")-Id. PFGE revealed heterogeneous patterns with no dominant clone. The results demonstrated that AME are common in aminoglycoside-resistant enterococci isolated in Kuwait. However, the absence of a dominant clone suggests that they acquired high-level aminoglycoside independently.

Point surveillance of Staphylococcus aureus carriage among medical staff in Infectious Diseases Hospital, Kuwait.

OBJECTIVE: To investigate the carriage of Staphylococcus aureus among doctors and nurses in Infectious Diseases Hospital, Kuwait, following the detection of 3 cases of methicillin-resistant S. aureus (MRSA). MATERIALS AND METHODS: A total of 260 nasal and throat swabs were obtained from 19 doctors and 111 nurses and cultured for the carriage of S. aureus. Forty-three S. aureus were identified based on their growth characteristics on mannitol-salt agar, catalase and tube coagulase and DNA hydrolysis. The isolates were tested for susceptibility to antibacterial agents and typed by phage typing; plasmid analysis and pulsed-field electrophoresis were carried out to determine their relatedness. RESULTS: Of the 19 doctors, 4 (21%) had nasal carriage while only 1 of them had a throat carriage. Sixteen (14.4%) nurses carried S. aureus in their noses and 20 (18%) in their throats. The combined nasal carriage rate for both doctors and nurses was 15.8%, and combined throat carriage was 16.6%. None of them carried MRSA. The isolates were resistant to penicillin G (90%), tetracycline (23.3%), erythromycin (9.3%) and cadmium (100%). Typing of the isolates showed a variety of phage types, plasmid and pulsed-field gel electrophoresis patterns. DISCUSSION: None of the doctors or nurses carried MRSA. Typing of the methicillin-susceptible strains that they carried demonstrated that the S. aureus were different, indicating an absence of a dominant clone capable of spreading. It is important to maintain a low carriage of S. aureus among health-care workers.

A chromosomal location of the mupA gene in Staphylococcus aureus expressing high-level mupirocin resistance.

OBJECTIVES: To investigate the genetic location of the mupA gene in high-level mupirocin-resistant Staphylococcus aureus isolates. MATERIALS AND METHODS: Antibiotic resistance was detected by disc diffusion. The Etest was used to determine mupirocin MIC. The presence of mupA was detected by PCR using specific primers. Curing, transfer experiments, pulsed-field gel electrophoresis (PFGE) and DNA hybridization were used to study the genetic location of mupA. RESULTS: The isolates had mupirocin MICs > 1024 mg/L and were resistant to methicillin, gentamicin, kanamycin, streptomycin, erythromycin, tetracycline, ciprofloxacin, cadmium acetate, propamidine isethionate and ethidium bromide. They carried two plasmids of approximately 26 and 2.8 kb. Curing and transfer experiments demonstrated that the 26 kb plasmid encoded resistance to cadmium acetate, propamidine isethionate and ethidium bromide. Loss of mupirocin resistance corresponded to the loss of a 40 kb DNA fragment from a 175 kb SmaI chromosomal fragment. The mupA gene was detected only in the genomic DNA of the mupirocin-resistant strains and in their derivatives cured of the 26 kb plasmid. A labelled mupA probe hybridized to the 175 kb SmaI fragment only in the mupirocin-resistant isolates. CONCLUSION: The absence of mupA on any of the plasmids and its detection only in the chromosomal DNA of the parents and in their derivatives cured of the 26 kb plasmid strongly supports a chromosomal location for mupA in these isolates.

Synthesis and antibacterial activity of 5-substituted oxazolidinones.

A series of 5-substituted oxazolidinones with varying substitution at the 5-position of the oxazolidinone ring were synthesized and their in vitro antibacterial activity was evaluated. The compounds demonstrated potent to weak antibacterial activity. A novel compound (PH-027) demonstrated potent antibacterial activity, which is comparable to or better than those of linezolid and vancomycin against antibiotic-susceptible standard and clinically isolated resistant strains of gram-positive bacteria. Although the presence of the C-5-acetamidomethyl functionality at the C-5 position of the oxazolidinones has been widely claimed and reported as a structural requirement for optimal antimicrobial activity in the oxazolidinone class of compounds, our results from this work identified the C-5 triazole substitution as a new structural alternative for potent antibacterial activity in the oxazolidinone class.

Antibiotic resistance of enterococci isolated at a teaching hospital in Kuwait.

Enterococci isolated in a teaching hospital were studied for their resistance to different antibiotics. Minimum inhibitory concentrations to high-level aminoglycosides and glycopeptide antibiotics were determined by agar dilution and E-test methods respectively. Genes encoding aminoglycoside-modifying enzymes were detected by the polymerase chain reaction (PCR). 195 enterococci were isolated from urines (54.3%), wounds (16.4%), blood (10.2%), and miscellaneous sources (18.9%). They consisted of E. faecalis (88.7%), E. faecium (9.2%), E. casseliflavus (1.5%) and E. bovis (0.5%). None of the enterococci produced penicillinase but 3.5% of them were resistant to ampicillin. They were also resistant to high-level gentamicin (15.9%), kanamycin (22.0%), streptomycin (21.0%), tetracycline (65.1%), erythromycin (62.6%), ciprofloxacin (36.1%), chloramphenicol (26.1%), vancomycin (3.0%) and teicoplanin (2.0%). Most of the high-level aminoglycoside-resistant isolates contained genes coding the bifunctional aminoglycoside modifying enzymes AAC(6')-APH(2"), APH(3') and ANT(6') but not the ANT(4') enzyme. The results demonstrated a low prevalence of vancomycin resistance among Enterococci in this hospital.

Genetic analysis of methicillin-resistant Staphylococcus aureus expressing high- and low-level mupirocin resistance.

Clinical strains of methicillin-resistant Staphylococcus aureus (MRSA) expressing high- and low-level mupirocin resistance were studied to determine the genetic location of mupirocin and other resistance determinants. Mupirocin resistance was confirmed by MIC determination with E-test strips. Curing and transfer experiments were used to establish the genetic location of the resistance determinants and the PCR with mupA-specific primers was used to detect the presence of mupA genes. High-level mupirocin-resistant isolates had MICs >1024 mg/L, whereas the low-level resistant isolates had MICs of 32-128 mg/L. The isolates carried plasmids ranging from 2.8 to 38 kb in size. All of them carried 26- and 3.0-kb plasmids, but only the high-level mupirocin-resistant isolates carried a 38-kb plasmid. Curing and transfer experiments revealed that the 26-kb plasmid encoded resistance to cadmium, mercuric chloride, propamidine isethionate and ethidium bromide and the 38-kb plasmid was a conjugative plasmid encoding high-level mupirocin resistance. One isolate, IBN287, carried both plasmid-borne high-level and chromosomal low-level mupirocin resistance. The mupA gene was detected on the 38-kb plasmid DNA but not in the genomic DNA of the low-level mupirocin-resistant isolates. The genomic DNA of strain IBN287 cured of the 38-kb mupirocin resistance plasmid did not contain mupA. The results suggest that different genes encoded low- and high-level mupirocin resistance in these isolates.

The spread of a mupirocin-resistant/methicillin-resistant Staphylococcus aureus clone in Kuwait hospitals.

High-level mupirocin- and methicillin-resistant Staphylococcus aureus (MRSA) isolated from five hospitals in Kuwait were studied by pulsed-field gel electrophoresis (PFGE) to determine their relatedness to one another and to high-level mupirocin-resistant MRSA isolated previously in a Burns Unit. The genetic location of mupirocin resistance determinant was also determined. All of the isolates were resistant to gentamicin, kanamycin, streptomycin, tetracycline and cadmium, and contained plasmids of 38, 26 and 2.8 kb. Two isolates contained additional 4.4-kb plasmids. Transfer experiments demonstrated that the 38-kb plasmid encoded high-level mupirocin resistance and the 4.4-kb plasmid encoded chloramphenicol resistance. PFGE typing of representative isolates from the five hospitals demonstrated that the majority of them had identical or closely related pulsed-field patterns suggesting that they had a common origin. However, they differed from high-level mupirocin-resistant MRSA isolated previously in the Burns Unit in their resistance and pulsed-field patterns. Whereas the majority of the previous isolates were susceptible to ciprofloxacin and resistant to trimethoprim and chloramphenicol, the majority of the current isolates were susceptible to trimethoprim and chloramphenicol, and resistant to ciprofloxacin. Only one of the current isolates had identical pulsed-field pattern to the majority of isolates obtained previously in the Burns Unit. The results suggested that a previously dominant clone of high-level mupirocin-resistant MRSA has been replaced in the Burns Unit by a new clone, which also spread in four other hospitals.

Persistence of a clone of methicillin-resistant Staphylococcus aureus in a burns unit.

A total of 128 MRSA isolates from a burns unit in 1992 and 1997 was studied by resistotyping, plasmid analysis and pulsed-field gel electrophoresis (PFGE) of SmaI-digested chromosomal DNA to ascertain whether a clone of MRSA had persisted in the unit or whether different clones had been introduced at different times. All the MRSA isolates produced beta-lactamase and had high MICs to methicillin (>256 mg/L). All were resistant to tetracycline, kanamycin, cadmium acetate and mercuric chloride. Most were resistant to gentamicin, neomycin, erythromycin, chloramphenicol, trimethoprim, ciprofloxacin, propamidine isethionate and ethidium bromide, and were susceptible to minocycline, vancomycin and teicoplanin. None of the 1992 isolates was resistant to mupirocin, but 56% and 19% of the 1997 isolates expressed high- and low-level mupirocin resistance, respectively. Many of the 1997 isolates had acquired a 38-kb plasmid encoding high-level mupirocin resistance. The 1992 isolates had two main PFGE patterns; 82% of them belonged to PFGE pattern 1. The 1997 isolates had PFGE pattern 1, the same as the majority of the 1992 isolates. All MRSA isolates from both years carried the mecA gene in the same SmaI fragment. These findings demonstrated that a clone of MRSA that was prevalentin the burns unit in 1992 had persisted and became the predominant clone in 1997.