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PubMed1999

Identification and medical importance of coagulase-negative staphylococci species.

A total of 126 coagulase-negative staphylococci strains (CNS) were isolated from blood samples and from the intravenous catheters and cerebrospinal fluid of 103 patients admitted to the University Hospital of Ribeirão Preto. Staphylococcus epidermidis (68.2%), S. haemolyticus (11.1%) and S. hominis (3.2%) were the most frequent species. The last two CNS showed greater resistance to antimicrobial agents than S. epidermidis. CNS were the agents of infection in 10. 7% of the patients and the agents of intravenous catheter colonization in 18.4% of the cases.

Bacteremia↗

PubMed2003

Five-test simple scheme for species-level identification of clinically significant coagulase-negative staphylococci.

A working scheme developed in our laboratory for identification (by species group and species) of coagulase-negative staphylococci (CNS) was evaluated with 201 consecutive isolates and then validated by using the reference method of Kloos and Schleifer (W. E. Kloos and K. H. Schleifer, J. Clin. Microbiol. 1:82-88, 1975). This five-test simple scheme (referred to here as the simple scheme) combines the novobiocin susceptibility test with tests for urease, pyrrolidonyl arylamidase, ornithine decarboxylase, and aerobic acid from mannose. The addition of one or two tests within a particular species group could then positively identify the isolate. Two commercial systems, Staph-Zym (Rosco) and API-Staph (bioMérieux), along with results obtained by using Rosco diagnostic tablets (nongrowth tests), were also compared with the reference method. One isolate could not be identified even by the reference method. Of the remaining 200 strains, 191 (95.5%) strains were correctly identified with Staph-Zym and 171 strains (85.5%) were correctly identified with API-Staph. The most frequent clinical CNS species isolated were Staphylococcus epidermidis (50.5%), S. haemolyticus (18.5%), S. saprophyticus subsp. saprophyticus (16.0%), S. lugdunensis (6.0%), and S. warneri (2.5%). The simple scheme validated with the reference method has demonstrated an excellent correlation in the identification of the three most frequent species isolated: S. epidermidis, S. haemolyticus, and S. saprophyticus subsp. saprophyticus. With the simple scheme, identification of CNS was possible within 24 h after the enzymatic tests were used, whereas up to 72 h is necessary for the growth tests. This methodology would be very useful in any clinical microbiology laboratory for the presumptive identification of CNS species groups and species.

Bacterial Typing Techniques↗

PubMed1984

Characterization of coagulase-negative staphylococci from patients with a malignancy compared with those from other sources.

170 strains of coagulase-negative staphylococci isolated from different patient groups have been characterized: 74% were Staphylococcus epidermidis, 18% S. haemolyticus and 7% S. simulans. There were no differences in rates of isolation of the various species from patients with and without a malignancy. Phage typing was not a useful epidemiological tool, but biotyping was possible using the API Staph system. Slime production, a characteristic said to predispose to colonization of catheters, was found to be most common in S. epidermidis strains.

Bacteriophage Typing↗

PubMed1994

Crystal violet reactions of coagulase negative staphylococci.

Twenty four reference strains and 112 clinical isolates of coagulase negative staphylococci (CNS) were examined for their reactions in the crystal violet test. Some species gave a white reaction and others a purple reaction. Results were consistent and reproducible and each species gave only one pattern of crystal violet reaction. Within the limited variety of species represented in the clinical isolates, Staphylococcus saprophyticus and S haemolyticus gave crystal violet purple reactions, in contrast to S epidermidis, which always gave a white reaction. Investigations suggested that the mechanism of the crystal violet test in S haemolyticus may be similar to that previously described in S aureus. Further work is needed to characterise the ability of crystal violet to modify S epidermidis and other central nervous system species. The crystal violet reaction, which has strong associations with invasiveness, phage group susceptibilities, colonisation persistence abilities, and nosocomial origin in S aureus may also be useful in studies of CNS disease.

Coagulase↗

PubMed1992

The epidemiology of ciprofloxacin resistance in coagulase-negative staphylococci in CAPD patients.

Ciprofloxacin was used as empirical therapy for peritonitis in patients receiving continuous ambulatory peritoneal dialysis (CAPD) for 26 months, providing an opportunity to study the epidemiology of ciprofloxacin resistance amongst coagulase-negative staphylococci (CNS). Swabs were collected from the CAPD patients, staff, and clinic environment before, during and after the time this antibiotic was prescribed. Clinical isolates were also studied, and records kept of patient hospital attendance. Ciprofloxacin-resistant staphylococci were typed by antibiogram, biotype, plasmid profile, SDS-PAGE, and immunoblotting. No resistant strains were detected before the use of ciprofloxacin. During its use 30% of patients became skin carriers, and resistant strains caused 8% of peritonitis episodes in 7% of patients (38% of carriers). Resistant strains were isolated from the environment, but never from attending members of staff. A total of 208 resistant isolates with MIC's between 8 and 128 mg/l was collected and identified as Staphylococcus epidermidis or S. haemolyticus. Sixteen strain types were distinguished. There was epidemiological evidence for selection of resistant strains derived from the host commensal flora and also for cross-colonization, and cross-infection. Carriage of resistant strains fell to 15% of patients, 6 months after the use of ciprofloxacin had ceased.

Air Microbiology↗

PubMed1997

Molecular typing of Staphylococcus epidermidis and other CNS with repetitive element sequence-based PCR.

The chromosomal distribution of the repetitive DNA sequence found in Mycoplasma pneumoniae (REP-MP2) provides an ideal target for detecting DNA fragment patterns specific to individual Staphylococcus epidermidis and S. haemolyticus strains. A REP-MP2 sequence-based PCR (rep-PCR) was developed and applied to CNS isolates. We identified a 450 bp genomic DNA fragment which was common and specific to S. epidermidis isolates and not found in other CNS. In addition, S. epidermidis isolates showed several bands that could be grouped into 14 different fragment patterns. Similarly, S. haemolyticus isolates were classified into 10 groups. Significant correlations between the typing patterns of S. epidermidis and resistance to oxacillin (P< 0.05), gentamicin (P< 0.01), erythromycin (P< 0.02), and sulfamethoxazole-trimethoprim (P< 0.001) were found. The rep-PCR method is a rapid and reproducible discriminatory means for molecular typing of S. epidermidis and other CNS.

Anti-Bacterial Agents↗

PubMed1985

Antimicrobial resistance in coagulase-negative staphylococci.

Patterns of resistance to antimicrobial agents were studied in 193 strains of coagulase-negative staphylococci isolated from hospital patients. Strains isolated from patients with malignant disease were significantly more often resistant to sulphonamide, trimethoprim, gentamicin and methicillin than were strains from other sources. Susceptibility to various beta-lactam antibiotics and aminoglycosides was investigated in members of the two most frequent species: Staphylococcus epidermidis and S. haemolyticus. S. haemolyticus strains were not only more often resistant to methicillin than S. epidermidis strains (respectively 81% and 17%) but they were more highly resistant (mean MICs respectively 85 and 19 mg/L). Methicillin-resistant S. haemolyticus strains were highly resistant to nine other beta-lactam antibiotics, whereas methicillin-resistant S. epidermidis strains showed both lower levels and a narrower spectrum of cross-resistance. Resistance to methicillin in members of both species was "heterogeneous", i.e., only a minority of cells in a culture showed significant resistance. Almost all gentamicin-resistant strains were sensitive to netilmicin and amikacin; rifampicin, vancomycin and teicoplanin were also highly active in vitro.

Aminoglycosides↗

PubMed1990

Sequential analysis of staphylococcal colonization of body surfaces of patients undergoing vascular surgery.

Slime-producing coagulase-negative staphylococci are pathogens in vascular surgery by virtue of their ability to adhere to and persist on prosthetic graft material. Inguinal and abdominal skin sites were cultured in 41 patients upon hospitalization, and slime production and antimicrobial susceptibility were assessed in all recovered staphylococcal isolates. Twenty-one patients eventually underwent lower-extremity revascularization. In the operative population, cultures were also obtained on the day of surgery and fifth postoperative day. All 21 patients received perioperative cefazolin. Of 327 coagulase-negative staphylococci recovered, Staphylococcus epidermidis (47%), S. haemolyticus (21%), and S. hominis (10%) were the predominant isolates. Slime-producing coagulase-negative staphylococci were recovered from 17 of 21 patients at admission but only from 8 of 21 patients on day 5 postoperation (P less than 0.05). S. epidermidis isolates demonstrated increasing multiple resistance from admission to 5 days postoperation to methicillin, gentamicin, clindamycin, erythromycin, and trimethoprim-sulfamethoxazole (P less than 0.05). All coagulase-negative staphylococcal isolates were susceptible to ciprofloxacin and vancomycin. Slime-producing capability was not associated with increased methicillin resistance for the recovered isolates. The data demonstrate that patients enter the hospital colonized with slime-producing strains of coagulase-negative staphylococci and that during hospitalization the staphylococcal skin burden shifts from a predominately susceptible to a resistant microbial population, which may enhance the importance of slime production as a risk factor in lower-extremity revascularization.

Blood Vessel Prosthesis↗

PubMed1991

Clinical results of a multicenter study with sulbactam/ampicillin for the treatment of patients with lower respiratory and urinary tract infections.

A total of 124 patients with lower respiratory tract (44) or urinary tract infections (80) were enrolled in an open, multicenter study to evaluate the efficacy and tolerability of sulbactam/ampicillin, administered at the dosage of 3 g/die by intramuscular route. Pretreatment pathogens from patients with lower respiratory tract infections included: Streptococcus alpha-haemolyticus in 8 cases, Streptococcus beta-haemolyticus in 2 cases, Staphylococcus albus in 7 cases, Haemophilus influenzae in 7 cases, Staphylococcus aureus in 6 cases, Klebsiella oxytoca in 5 cases, Staphylococcus epidermidis in 3 cases, Streptococcus pneumoniae in 3 cases, Escherichia coli in 2 cases; in one subject (2.75%), no microorganisms were isolated. In vitro, 36 isolates (84%) were sensitive to SA and 7 (16%) were resistant. At the end of therapy, all the causative pathogens sensitive to sulbactam/ampicillin were eliminated. In patients with urinary tract infections, pretreatment pathogens were: E. coli in 40 cases, S. albus in 16 cases, Proteus mirabilis in 8 cases, Enterobacter agglomerans in 6 cases, Proteus vulgaris in 3 cases, Streptococcus faecalis in 3 cases, Streptococcus liquefaciens in 2 cases, Pseudomonas aeruginosa in 2 cases. In vitro, 64 isolates (80%) were sensitive to sulbactam/ampicillin and 16 (20%) were resistant. At the end of therapy, 63 out of the 64 pathogens sensitive to sulbactam/ampicillin were eliminated; in one case the therapy was interrupted due to adverse effect. Clinical efficacy: in subjects with lower respiratory tract infections, sulbactam/ampicillin cured 32 patients (72.72%) and ameliorated the clinical status of 8 patients (18.18%); efficacy rate: 90.9%).(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

PubMed1993

Octapeptide derivatives of teicoplanin antibiotics.

A series of octapeptide derivatives of teicoplanin-A2 component 2 (CTA/2), its aglycone (TD), and the L-lysyl derivatives of an amide of CTA/2 and TD, were prepared by condensation of the terminal amino group with N-hydroxysuccinimidyl esters of tert-butyloxycarbonyl (BOC) L- and D-amino acids, followed by acidic (TFA) removal of the BOC protecting function. The antimicrobial properties of these compounds were compared with those of the corresponding unmodified antibiotics and their N15-acetyl derivatives. The most active derivatives were the octapeptides with N-terminal glycine or lysine whose in vitro activity was comparable to that of the parent teicoplanins. The glycinyl and lysyl derivatives of CTA/2 showed better activity than CTA/2 against clinical isolates of Staphylococcus epidermidis and S. haemolyticus for which teicoplanin MICs were relatively high. No significant difference in their antibacterial activity was observed between octapeptides containing L- or D-lysine.

Anti-Bacterial Agents↗

PubMedDate not supplied

[Quality control of erythrocyte concentrates in 6 transfusion centers in Croatia].

Quality control of red cell concentrates (RCC) was performed in six Blood Transfusion Centres in Croatia. Between 1.5 and 3.7% of the produced or/and used RCC were examined. Large differences in volume and haematocrit were found between red cell concentrates. 63.2% of concentrates had a volume larger than 225 ml. The mean haematocrit was 65.5%, and a haematocrit above 65% was observed in 51.6% of RCC. From 3.5% of RCC bacteria were isolated (gram positive coccus, Staphylococcus albus, Staphylococcus epidermidis and Streptococcus haemolyticus). The quality of RCC was not found to be satisfactory. In order to improve their quality procedures for their production, standards and legal requirements have to be established.

Blood Transfusion↗

PubMed2006

Clinical significance of coagulase-negative staphylococci recovered from nonsterile sites.

Laboratory criteria were used to select coagulase-negative staphylococci isolated from nonsterile body sites for further investigation. Fifty-seven percent of the study isolates were clinically significant, predominantly causing community-acquired soft tissue infections. There were species-related differences in virulence, with 91% of Staphylococcus lugdunensis isolates clinically significant compared with 11% of S. haemolyticus isolates.

Algorithms↗

PubMed1992

Survey of methicillin-resistant clinical strains of coagulase-negative staphylococci for mecA gene distribution.

A total number of 125 methicillin-resistant (MIC, greater than or equal to 16) coagulase-negative Staphylococcus strains isolated in Japan were surveyed for the distribution of the mecA gene, the structural gene for penicillin-binding protein 2', which is the causative genetic element for the intrinsic resistance of methicillin-resistant Staphylococcus aureus. Screening with colony hybridization by using a cloned mecA gene probe revealed that 121 strains (96.8%) belonging to the nine coagulase-negative Staphylococcus species (S. epidermidis, S. haemolyticus, S. saprophyticus, S. sciuri, S. simulans, S. hominis, S. capitis, S. warneri, and S. caprae) carried mecA in their genome, indicating wide distribution of the gene among coagulase-negative Staphylococcus species. Most (93.4%) of the mecA-carrying strains were producers of penicillinase. Four strains, including two S. haemolyticus and two S. saprophyticus strains, did not carry mecA in spite of their resistance to methicillin. One of them was of low-level resistance (MIC, 16), but three of them had moderate- to high-level resistance to methicillin (MIC, 64). Analysis of gel electrophoretic banding patterns of penicillin-binding proteins of these strains showed absence of penicillin-binding protein 2' but some alterations in signal intensities of the other penicillin-binding proteins. The result indicated that about 3% of methicillin-resistant coagulase-negative staphylococci in these hospitals had a resistance mechanism different from that associated with the production of penicillin-binding protein 2', as has been reported in the case of a borderline methicillin-resistant strain of S. aureus.

Bacterial Proteins↗

PubMed1997

Identification of Staphylococcus species and subspecies by the chaperonin 60 gene identification method and reverse checkerboard hybridization.

A previous study (S. H. Goh et al., J. Clin. Microbiol. 34:818-823, 1996) demonstrated that a 600-bp region of the chaperonin 60 (Cpn60) genes from various bacterial isolates could be amplified by PCR with a pair of degenerate primers and that the products could be used as species-specific probes for Staphylococcus aureus, S. epidermidis, S. haemolyticus, S. lugdunensis, S. saprophyticus, and S. schleiferi. To further validate the utility of bacterial Cpn60 genes as universal targets for bacterial identification (ID), reverse checkerboard chemiluminescent hybridization experiments were performed with DNA probes from 34 different Staphylococcus species and subspecies. With the exception of probes from the Cpn60 genes of S. intermedius and S. delphini, which cross hybridized, all were species specific. Two subspecies of both S. capitis and S. cohnii were differentiated from one another, while DNAs from the two S. schleiferi subspecies cross hybridized. When 40 known Staphylococcus isolates were tested in a blind experiment by the Cpn60 gene method, 36 strains, representing six species and one subspecies (S. sciuri, S. caseolyticus, S. hominis, S. warneri, S. hyicus, S. haemolyticus, and S. capitis subsp. ureolyticus), were correctly identified. DNA from the four remaining isolates, known to be S. hyicus bovine strains, failed to hybridize to DNA from the S. hyicus target strain or any other Staphylococcus species. However, DNAs from these S. hyicus isolates did cross hybridize with each other. New DNA sequence data and evidence from previous studies suggest some genetic divergence between the two groups of S. hyicus isolates. Our results demonstrate that this Cpn60 gene-based ID method has the potential to be a basic method for bacterial ID. Studies are in progress to further validate the utility of this Cpn60 gene system for ID of Staphylococcus and other genera, including those of slow-growing microorganisms.

Animals↗

PubMed1990

Percentages and distributions of teicoplanin- and vancomycin-resistant strains among coagulase-negative staphylococci.

The activities of teicoplanin and vancomycin against 362 coagulase-negative staphylococci were determined by an agar dilution method. At the 4- and 32-micrograms/ml breakpoint levels of the National Committee for Clinical Laboratory Standards, 23.2% of the strains were intermediate and 1.7% were resistant to teicoplanin, in contrast to less than 0.3% intermediate to vancomycin. Resistant strains belonged to the species Staphylococcus epidermidis (74%) and S. haemolyticus (19%).

Coagulase↗

PubMed1991

Serological typing of coagulase-negative staphylococci using monoclonal antibodies.

This investigation was to determine whether monoclonal antibodies (Mabs) could be used to differentiate coagulase-negative staphylococci (C-NS) at species and strain level. Mabs were produced to four Staphylococcus epidermidis strains, two S. haemolyticus strains, one S. saprophyticus strain and one S. warneri strain. A panel of nine antibodies was tested for species and strain specificity against five type strains and 65 clinical isolates of C-NS by enzyme-linked immunosorbent assay (ELISA). Species specificity was found with Mab D150 produced to one S. haemolyticus strain. Using Mab D150 and Mab D198 in conjunction, identification of 90% of S. haemolyticus isolates to species level was achieved. S. saprophyticus Mab K84 reacted with most other strains of C-NS tested but only three S. haemolyticus strains (16%). This finding provides further evidence that S. haemolyticus possesses different surface determinants to other C-NS which could form the basis of a typing scheme for S. haemolyticus using Mabs D150, D198 and K84.

Antibodies, Monoclonal↗

PubMed2000

Nationwide German multicenter study on prevalence of antibiotic resistance in staphylococcal bloodstream isolates and comparative in vitro activities of quinupristin-dalfopristin.

Antibiotic-resistant gram-positive bacteria have become an increasing problem in the last two decades. In order to evaluate the prevalence of antibiotic resistance in staphylococcal bloodstream isolates in Germany, 2,042 staphylococci collected in 21 tertiary-care hospitals were investigated during a 3-year period (March 1996 to March 1999). Altogether, 1,448 S. aureus isolates and 594 coagulase-negative staphylococci (CoNS) that comprised 13 different species were included. Furthermore, the antistaphylococcal activities of quinupristin-dalfopristin were compared with those of eight other compounds by the broth microdilution method. The rates of oxacillin resistance in Staphylococcus aureus, S. epidermidis, S. haemolyticus, and other CoNS were 13.5, 69, 90, and 34%, respectively. In oxacillin-resistant strains high rates of resistance (up to 100%) to erythromycin, clindamycin, ciprofloxacin, and gentamicin were also observed. However, no strain appeared to be resistant to vancomycin or quinupristin-dalfopristin. The streptogramin combination exhibited excellent in vitro activity against all staphylococcal species tested, regardless of the patterns of resistance to other drug classes. In terms of MICs at which 90% of the isolates are inhibited, quinupristin-dalfopristin was 2 times more active against S. aureus isolates, 4 to 16 times more active against S. haemolyticus, and 8 to 32 times more active against S. epidermidis than vancomycin or teicoplanin.

Anti-Bacterial Agents↗

PubMed1998

Coagulase-negative staphylococci isolated from various skin lesions.

We isolated 162 coagulase-negative staphylococci (48: from infection, 114: from colonization) from various skin diseases between January, 1995, and January, 1998. From eighteen infected cysts, 10 Staphylococcus epidermidis strains, 3 S. capitis strains, 2 S. hominis strains, 2 S. auricularis strains, and one S. saprophyticus strain were individually detected. Similarly, from ten folliculitis lesions, 6 S. epidermidis strains, 2 S. capitis strains, and 2 S. hominis strains, and from five furuncle lesions, 3 S. lugdunensis strains, one S. epidermidis strains, and one S. hominis strain were detected, respectively. Four abscesses with mild inflammatory signs were localized on the scalp; S. epidermidis strains alone were detected from them. From two felons, one S. lugdunensis strain and one S. haemolyticus strain were detected, respectively. Staphylococcus epidermidis and S. lugdunensis strains seems to be more frequently associated with skin suppurative lesions than other strains. Staphylococcus hominis strains and S. capitis strains were suggested to be potential pathogens in the initiation of suppuration in various purulent skin lesions. Among the 28 S. epidermidis strains, 13 (46.4%) were methicillin-resistant (oxacillin, minimum inhibitory concentration > or = 4 micrograms/ml). Twelve (29.3%) out of the other 41 coagulase-negative staphylococci were methicillin-resistant. Coagulase-positive and -negative staphylococci showed no differences in susceptibility tests against various antistaphylococcal agents.

Anti-Bacterial Agents↗