A trial of the use of mupirocin in the nasal carriage of Staphylococcus aureus in medical personnel]

Many hospital-acquired purulent diseases and wound infections are due to multiresistant hospital strains of Staphylococcus aureus. The role of S. aureus nasal carriage in development of wound infections due to autoinfection is confirmed. Not only inpatients but also hospital staff can be highly colonized with coagulase positive staphylococci. The S. aureus persistence in hospital personnel results in distribution of the microorganisms in the environment. Therefore, detection of S. aureus carriers without signs of the infection among the hospital personnel and eradication of the pathogen make it possible to control outbreaks of S. aureus infection in hospitals. Clinical efficacy of nasal ointment of mupirocin in the treatment of S. aureus carriers among the intensive care personnel of the N. N.Blokhin Cancer Research Center was evaluated. S. aureus nasal carriage was diagnosed in 17 (26 per cent) out of 65 persons. All the isolates were susceptible to oxacillin. 5-7 days after discontinuation of the mupirocin nasal ointment use eradication of S. aureus was stated in 100 per cent of the cases. The effect was still observed in 94 per cent of the cases in 1 month, in 76 per cent of the cases in 5-6 months and in 60 per cent of the cases in 8-9 months. It is believed that mupirocin nasal ointment (Bactroban) is convenient to use, low toxic and highly active in the treatment of persons with S. aureus nasal carriage.     

Molecular epidemiology of infections caused by methicillin-resistant staphylococci

The review deals with the periodicity of the spread of methicillin-resistant S. aureus (MRSA) strains during the last 40 years, the mechanism of their resistance to methicillin and other beta-lactamic antibiotics, the genetic control of methicillin resistance, the genome organization of mec DNA and its possible cause, as well as the organization of epidemiological surveillance on MSRA in hospitals. The problem of changes in the epidemiology of staphylococcal infections due to the appearance of MRSA in the absence of contacts with carriers, treatment with antibiotics or stay in a hospital is discussed. The concern of public health authorities in connection with the emergence of MRSA strains, moderately resistant or resistant to vancomycin, is also discussed. The most promising programs of the MRSA study, as well as the optimum programs introduced in economically developed counties for the control of hospital infections caused by MRSA, are considered.     

Mechanism of the heterogeneity of a staphylococcal population with respect to methicillin resistance]

The study of the staphylococcal population heterogeneity with respect to methicillin resistance by 2 methods revealed different numbers of the resistant cells in the population. Thus, when the microbial suspension was plated on an agarized medium with methicillin (50 gamma/ml), only 0.0007--0.0005 per cent of the resistant cells were found. When the colonies were replicated from a medium without methicillin to a medium containing methicillin (50 gamma/ml), 84.3--97.3 per cent of the resistant microbial cells were found in the population of the same strains. The main mechanism in the heterogeneity of the staphylococcal population with respect to methicillin resistance was impairement of the phenotype manifestation of the antibiotic resistance under definite conditions.     

Molecular epidemiology of clinical and carrier strains of methicillin resistant Staphylococcus aureus (MRSA) in the hospital settings of north India

Background

The study was conducted between 2000 and 2003 on 750 human subjects, yielding 850 strains of staphylococci from clinical specimens (575), nasal cultures of hospitalized patients (100) and eye & nasal sources of hospital workers (50 & 125 respectively) in order to determine their epidemiology, acquisition and dissemination of resistance genes.

Methods

Organisms from clinical samples were isolated, cultured and identified as per the standard routine procedures. Susceptibility was measured by the agar diffusion method, as recommended by the Nat ional Committee for Clinical Laboratory Standards (NCCLS). The modified method of Birnboin and Takahashi was used for isolation of plasmids from staphylococci. Pulsed-field gel electrophoresis (PFGE) typing of clinical and carrier Methicillin resistant Staphylococcus aureus (MRSA) strains isolated during our study was performed as described previously.

Results

It was shown that 35.1% of Staphylococcus aureus and 22.5% of coagulase-negative staphylococcal isolates were resistant to methicillin. Highest percentage of MRSA (35.5%) was found in pus specimens (n = 151). The multiple drug resistance of all MRSA (n = 180) and Methicillin resistant Coagulase-negative Staphylococcus aureus (MRCNS) (n = 76) isolates was detected. In case of both methicillin-resistant as well as methicillin-sensitive Saphylococcal isolates zero resistance was found to vancomycin where as highest resistance was found to penicillin G followed by ampicillin. It was shown that the major reservoir of methicillin resistant staphylococci in hospitals are colonized/infected inpatients and colonized hospital workers, with carriers at risk for developing endogenous infection or transmitting infection to health care workers and patients. The results were confirmed by molecular typing using PFGE by SmaI-digestion. It was shown that the resistant markers G and T got transferred from clinical S. aureus (JS-105) to carrier S. aureus (JN-49) and the ciprofloxacin (Cf) and erythromycin (E) resistance seemed to be chromosomal mediated. In one of the experiments, plasmid pJMR1O from Staphylococcus aureus coding for ampicillin (A), gentamicin (G) and amikacin (Ak) resistance was transformed into Escherichia coli. The minimal inhibitory concentrations (MICs) for A and G were lower in E. coli than in S. aureus. However, the MIC for Ak was higher in E. coli transformants than in S. aureus.

Conclusion

There is a progressive increase in MRSA prevalence and multi-drug resistance in staphylococci. Vancomycin is still the drug of choice for MRSA infections. The major reservoir of methicillin resistant staphylococci in hospitals is colonized/infected inpatients and colonized hospital workers. Resistance transfer from staphylococci to E. coli as well as from clinical to carrier staphylococci due to antibiotic stress seemed to be an alarming threat to antimicrobial chemotherapy.     

Nasal carriage of methicillin resistant Staphylococcus aureus and their antibiotic susceptibility patterns in children attending day-care centers

Nasal colonization with community acquired methicillin resistant Staphylococcus aureus (CA-MRSA) is being increasingly reported, especially in places where people are in close contact and in reduced hygiene, such as day-care centers. In this study we investigated the frequency of MRSA colonization and their antibiotic susceptibility patterns in 1-6 years old children of day-care centers in Hamadan, West of Iran.Five hundred nasal swabs were collected from children of 27 day-care centers that had no risk factors for colonization by S. aureus. The specimens were cultured for isolation of S. aureus by standard methods. Antimicrobial susceptibility testing was performed according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. For evaluation of the frequency of erythromycin induced clindamycin resistance, disk approximation test (D-test) was applied.Totally, 148 (29.6%) children were colonized by S. aureus. Out of 260 male, 94 (36.2%) and of 240 female, 54 (22.5%) cases were nasal carriers of S. aureus (P value = 0.001). Six (4.1%) of the 148 S. aureus isolated from children were MRSA strains. None of MRSA and methicillin susceptible S. aureus (MSSA) was resistant to vancomycin and clindamycin. Three of the 6 strains of MRSA and 7 (4.9%) of the 142 MSSA strains were resistant to erythromycin, and D-test was positive in all of them.We conclude that the rate of colonization by S. aureus is high in children attending day-care centers but colonization with MRSA is not common in our areas. Clindamycin or trimethoprim-sulfamethoxazol could be used in mild to moderataly severe diseases caused by CA-MRSA. However, if the CA-MRSA isolates are erythromycin resistant, D-test should be carried out for detection of inducible clindamycin resistance.     

Antibiotic sensitivity of Staphylococci populating the breast skin of nursing women

Sensitivity to penicillins, tetracycline, erythromycin and lincomycin was tested in 1513 staphylococcal strains isolated from the skin of various anatomic areas of the breast of nursing mothers. It was shown that 82.9, 78.4 and 33.9 per cent of the isolated cultures were highly sensitive to lincomycin, erythromycin and methicillin respectively. Sensitivity to tetracycline and benzylpenicillin was detected in 33.1 and 19.8 per cent of the cultures respectively. The study of the resistance spectra of 1343 strains resistant to certain antibiotics revealed that 498 cultures (37.1 per cent) were polyresistant. None of the tested antibiotics could be used for selective decontamination of the breast skin with respect to S. aureus in prophylaxis of lactic mastitis.     

The investigation of Staphylococcus aureus and coagulase-negative staphylococci nasal carriage among patients undergoing haemodialysis

The frequency of nasal staphylococcal colonization among haemodialysed patients was investigated. The swabs were collected in 1998 and 2004 from 28 and 43 patients, respectively.

Staphylococcus aureus colonization rates were 57.1% and 27.9% in 1998 and 2004, respectively. Twenty-six coagulase-negative staphylococci (CNS) isolates were cultured: S. epidermidis (21), S. lugdunensis (2), single S. haemolyticus, S. warneri, and S. capitits isolates. One S. aureus and 10 CNS isolates were methicillin resistant. The methicillin-resistant S. aureus (MRSA) was resistant to β-lactams, tetracycline, and harbored the pvl gene encoding the Panton-Valentine leukocidin.

The decrease in S. aureus colonization at 6-year interval was observed. The presence of the pvl gene and a favorable antibiotic susceptibility pattern of the MRSA suggest that the isolate was a member of community-acquired MRSA (CA-MRSA). Concluding, screening of haemodialysed patients for staphylococcal colonization accompanied by characterization of cultured isolates is important to understand its epidemiology and to develop infection prevention measures and treatment strategies.     

Reduced susceptibility to triclosan in methicillin-resistant Staphylococcus aureus

Susceptibility to triclosan in Staphylococcus aureus was determined. The study was carried out on 200 strains, including 100 resistant (MRSA) and 100 susceptibile (MSSA) to methicillin. The examined strains were isolated from varied clinical samples and patients in 18 medical centers, in majority from hospitals in the region of Gdansk. The susceptibility was estimated by the MIC (minimal inhibitory concentration) using dilution test in Mueller-Hinton agar. The antimicrobial resistance patterns were determined, including resistance to methicillin and mupirocin. The most of MRSA strains (62%) demonstrated reduced susceptibility to triclosan (MIC 2mg/L), while 93% of MSSA strains were highly sensitive to this antibacterial agent (MIC 0,031mg/L). The majority (66,1%) of MRSA strains with reduced susceptibility to triclosan demonstrated the same antimicrobial resistance pattern. Reduced susceptibility to triclosan was observed in 8 from 9 high - level mupirocin resistant strains, but the most of MRSA strains with reduced triclosan susceptibility (91,5%) were found among fusidic acid resistant strains.     

Production of toxic shock exotoxin by Staphylococcus aureus strains resistant and sensitive to antibiotics

Three hundred and ninety two strains of S. aureus isolated from bacteria carriers and patients with staphylococcal infections in different regions of the Soviet Union were investigated. 55.9 per cent of the isolates were able to produce exotoxin of toxic shock. No regular relation between resistance to definite antibiotics (tetracycline, chloramphenicol, benzylpenicillin, streptomycin, lincomycin, erythromycin, oleandomycin, gentamicin and methicillin) and the polyresistance range on the one hand and the ability to produce toxic shock exotoxin on the other hand was revealed.     

Donor activity of methicillin resistant staphylococci in transduction experiments]

Antibiotic resistance gene transmission from methicillin resistant strains of Staphylococcus aureus (MRSA), isolated in a burn care unit, was studied in transduction experiments with type phages 29, 52, 52A and in experiments with prophage induction. The results of the experiments demonstrated high donor activity of MRSA. Recombinants with different antibiotic resistance phenotypes were revealed, but there were no methicillin resistant staphylococci among them. Stability of cloramphenicol resistance gene transmission in the experiments on specific transduction with the prophage induction could be indicative of the prophage localization near the chloramphenicol resistance genes. Variety of the antibiotic resistance combinations in the transductants from the clinical strains of MRSA could prove heterogeneity of the strains even under conditions of one hospital.     

Antimicrobial resistance and presence of <Emphasis Type="Italic">ileS-2</Emphasis> gene encoding mupirocin resistance in clinical isolates of methicillin-resistant <Emphasis Type="Italic">Staphylococcus</Emphasis> sp.

Seventy-eight staphylococcal strains were isolated from surgical-site, blood-stream and other hospital-acquired infections. Eighteen isolates were determined as methicillin (MET)-resistant S. aureus (MRSA), while the remaining were MET-resistant coagulase-negative staphylococci (CoNS). Fifty percent of CoNS strains were multiresistant, while 10 % of isolates were resistant only to β-lactams. Clinical isolates of CoNS were generally more resistant to antimicrobial agents than S. aureus strains. Thirty-nine % of S. aureus strains were resistant only to β-lactams. None of the MRSA strains carried ileS-2 gene; this gene was found in two strains of S. epidermidis.     

Antibiotic resistant Staphylococcus aureus: a paradigm of adaptive power

Nothing documents better the spectacular adaptive capacity of Staphylococcus aureus than the response of this important human and animal pathogen to the introduction of antimicrobial agents into the clinical environment. The effectiveness of penicillin introduced in the early 1940s was virtually annulled within a decade because of the plasmid epidemics that spread the ss-lactamase gene through the entire species of S. aureus. In 1960 within one to two years of the introduction of penicillinase resistant ss-lactams (methicillin), methicillin resistant S. aureus (MRSA) strains were identified in clinical specimens. By the 1980s, epidemic clones of MRSA acquired multidrug resistant traits and spread worldwide to become one of the most important causative agents of hospital acquired infections. In the early 2000s, MRSA strains carrying the Tn1546 transposon-based enterococcal vancomycin resistant mechanism were identified in clinical specimens, bringing the specter of a totally resistant bacterial pathogen closer to reality. Then, in the late 1990s, just as effective hygienic and antibiotic use policies managed to bring down the frequency of MRSA in hospitals of several countries, MRSA strains began to show up in the community.     

Evaluation of BacLiteRapidMRSA, a rapid culture based screening test for the detection of ciprofloxacin and methicillin resistantS. aureus(MRSA) from screening swabs

Background  

Methicillin-resistantStaphylococcus aureus(MRSA) is a major nosocomial pathogen worldwide. The need for accurate and rapid screening methods to detect MRSA carriers has been clearly established. The performance of a novel assay, BacLiteRapidMRSA (Acolyte Biomedica, UK) for the rapid detection (5 h) and identification of hospital associated ciprofloxacin resistant strains of MRSA directly from nasal swab specimens was compared to that obtained by culture on Mannitol salt agar containing Oxacillin (MSAO) after 48 h incubation.     

The emergence and evolution of methicillin-resistant Staphylococcus aureus

Significant advances have been made in recent years in our understanding of how methicillin resistance is acquired by Staphylococcus aureus. Integration of a staphylococcal cassette chromosome mec (SCCmec) element into the chromosome converts drug-sensitive S. aureus into the notorious hospital pathogen methicilin-resistant S. aureus (MRSA), which is resistant to practically all beta-lactam antibiotics. SCCmec is a novel class of mobile genetic element that is composed of the mec gene complex encoding methicillin resistance and the ccr gene complex that encodes recombinases responsible for its mobility. These elements also carry various resistance genes for non-beta-lactam antibiotics. After acquiring an SCCmec element, MRSA undergoes several mutational events and evolves into the most difficult-to-treat pathogen in hospitals, against which all extant antibiotics including vancomycin are ineffective. Recent epidemiological data imply that MRSA has embarked on another evolutionary path as a community pathogen, as at least one novel SCCmec element seems to have been successful in converting S. aureus strains from the normal human flora into MRSA.     

Detection of methicillin-resistant <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> using double duplex real-time PCR and dye Syto 9

A screening method for methicillin-resistant Staphylococcus aureus (MRSA) using real-time polymerase chain reaction (PCR) and dye Syto 9 was developed and evaluated. The assay was based on the two duplex reactions run simultaneously. The detection reaction amplified staphylococcal cassette chromosome mec (SCCmec) right extremity sequences and S. aureus-specific 442-bp DNA (Sa442). The control reaction amplified S. aureus-specific nuclease gene nuc and a marker of methicillin resistance, mecA. The method was evaluated by analyzing 214 clinical S. aureus isolates yielding 98.7 % sensitivity, 100 % specificity, 100 % positive predictive value and 96.6 % negative predictive value for detection of MRSA. The detection limit was determined to be 15–80 genome copies per real-time PCR. It was able to discriminate between MRSA, methicillin resistant coagulase negative staphylococci and methicillin susceptible S. aureus (MSSA) isolates containing only small fragments of the right extremity of the SCCmec (MSSA revertants).     

Sensitivity to antibiotics and the disinfectant chloramine of Staphylococci isolated from healthy and sick persons

Five hundred and twenty seven strains of Staphylococcus aureus with massive contamination of at least 10(3) when estimated quantitatively were tested for their sensitivity to antibiotics and chloramine B. The staphylococcal strains were isolated from patients, air and stock of rooms in medical institutions, from medical personnel and healthy persons having no long-term contacts with hospital media i. e. from pregnant women and workers of confectionery plants. Among the isolates there were strains simultaneously sensitive to antibiotics and chloramine B (16.6 per cent), sensitive to antibiotics but resistant to chloramine B (5.5 per cent), resistant to antibiotics but sensitive to chloramine B (63.1 per cent) and resistant to antibiotics and chloramine B (15.3 per cent). Staphylococci resistant simultaneously to antibiotics and chloramine B were isolated from the persons of all the groups and from the air and stock of the rooms in the medical institutions. This showed the necessity of controlling sensitivity or staphylococci circulating in hospital media not only to antibiotics but also to disinfectants for providing more efficient prophylaxis of intrahospital infections.     

mecA基因PCR扩增法检测耐甲氧西林金黄色葡萄球菌

目的 应用mecA基因PCR扩增法检测耐甲氧西林金黄色葡萄球菌（methicillin resistant staphylococcus aureus,MRSA）。方法 临床分离的70株金黄色葡萄球菌,应用mecA基因PCR扩增法鉴定MRSA,并与苯唑西林纸片扩散法进行比较。结果 70株金黄色葡萄球菌用PCR扩增法和纸片扩散法有6株鉴定有差异,4株。mecA基因阳性而纸片扩散法鉴定为敏感,1株mecA基因阳性纸片扩散法鉴定为临界耐药,1株mecA基因阴性却表现为苯唑西林耐药,2种方法符合率为91．43％。结论 mecA基因PCR扩增法可以准确、快速判定MRSA,特别是对隐匿型或低水平耐药菌株的检出有重要的价值。     

Mutation in the C-Di-AMP Cyclase dacA Affects Fitness and Resistance of Methicillin Resistant Staphylococcus aureus

Faster growing and more virulent strains of methicillin resistant Staphylococcus aureus (MRSA) are increasingly displacing highly resistant MRSA. Elevated fitness in these MRSA is often accompanied by decreased and heterogeneous levels of methicillin resistance; however, the mechanisms for this phenomenon are not yet fully understood. Whole genome sequencing was used to investigate the genetic basis of this apparent correlation, in an isogenic MRSA strain pair that differed in methicillin resistance levels and fitness, with respect to growth rate. Sequencing revealed only one single nucleotide polymorphism (SNP) in the diadenylate cyclase gene dacA in the faster growing but less resistant strain. Diadenylate cyclases were recently discovered to synthesize the new second messenger cyclic diadenosine monophosphate (c-di-AMP). Introduction of this mutation into the highly resistant but slower growing strain reduced resistance and increased its growth rate, suggesting a direct connection between the dacA mutation and the phenotypic differences of these strains. Quantification of cellular c-di-AMP revealed that the dacA mutation decreased c-di-AMP levels resulting in reduced autolysis, increased salt tolerance and a reduction in the basal expression of the cell wall stress stimulon. These results indicate that c-di-AMP affects cell envelope-related signalling in S. aureus. The influence of c-di-AMP on growth rate and methicillin resistance in MRSA indicate that altering c-di-AMP levels could be a mechanism by which MRSA strains can increase their fitness levels by reducing their methicillin resistance levels.     

Comparison of methods for the detection of methicillin resistance in Staphylococcus aureus isolates from food products

AIMS: To compare several methods for detection of methicillin resistance in Staphylococcus aureus isolates from food. METHODS AND RESULTS: Two hundred S. aureus isolates from food of animal origin were screened for methicillin resistance by a PCR assay specific for the mecA gene, an oxacillin agar screen test and a cefoxitin disk diffusion test. Six out of 200 strains (3%) were found to be methicillin-resistant Staphylococcus aureus (MRSA) by PCR. The oxacillin agar screen test detected only one of the MRSA isolates (sensitivity of 16.7%) and mischaracterized three additional strains as MRSA (specificity of 98.45%). None of the MRSA strains was detected by the cefoxitin test (sensitivity of 0%), while 15 methicillin-susceptible S. aureus (MSSA) strains were misclassified as resistant (specificity of 92.3%). Fifteen MSSA strains displayed a beta-lactamase hyperproducer-like phenotype. The six MRSA (mecA-positive) strains resembled the characteristics of heteroresistant strains. CONCLUSIONS: As MRSA of animal origin may display atypical phenotypes, PCR appears to be more reliable for detection of methicillin resistance in animal strains. SIGNIFICANCE AND IMPACT OF THE STUDY: The study stresses the need for implementing the methods of screening S. aureus from food of animal origin for methicillin resistance.