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Multiplex PCR protocol for the diagnosis of staphylococcal infection.

We report the development of a multiplex PCR protocol for the diagnosis of staphylococcal infection. The protocol was designed to (i) detect any staphylococcal species to the exclusion of other bacterial pathogens (based on primers corresponding to Staphylococcus-specific regions of the 16S rRNA genes), (ii) distinguish between S. aureus and the coagulase-negative staphylococci (CNS) (based on amplification of the S. aureus-specific clfA gene), and (iii) provide an indication of the likelihood that the staphylococci present in the specimen are resistant to oxacillin (based on amplification of the mecA gene). The expected fragments were amplified from each of 60 staphylococcal isolates (13 oxacillin-resistant S. aureus isolates, 23 oxacillin-sensitive S. aureus isolates, 17 oxacillin-resistant CNS, and 7 oxacillin-sensitive CNS). No amplification products were observed with template DNA from nonstaphylococcal species, and the efficiency of amplification of staphylococcal targets was not adversely affected by the presence of DNA from other bacterial species in the same sample. The utility of the protocol for the analysis of clinical samples was verified by analysis of aliquots taken directly from BacT/Alert blood culture bottles. Of 77 blood cultures tested, only 7 yielded results inconsistent with those of conventional methods of diagnosis and susceptibility testing. Of those, one was identified as a CNS species by PCR and S. aureus by conventional methods. We also identified two isolates that were mecA positive but were oxacillin sensitive according to conventional methods. The other four samples failed to yield any amplification product even with a control set of primers corresponding to a conserved region of the eubacterial rRNA genes.

Blood↗

Vaccines and immunotherapy for staphylococcal infections.

Nosocomial or hospital-acquired infections are associated with prolonged hospitalizations and increased healthcare costs. Infections associated with surgical implants are becoming more difficult and more costly to manage, as they require repeated surgical procedures and a longer period of time to treat patients. Continued advances in the use of medical devices, an increase in the number of immunocompromised patients, and a steady rise in the prevalence of antibiotic-resistant organisms has renewed interest in the development of novel therapies that can be used to prevent and treat nosocomial infections. This review provides an overview of bacterial adhesins and focuses on novel immunological therapies developed to treat staphylococcal infections.

Adhesins, Bacterial↗

Diagnosis and management of staphylococcal infections of pacemakers and cardiac defibrillators.

Staphylococcal species, usually Staphylococcus aureus or Staphylococcus epidermidis, account for 70% to 95% of pacemaker and cardiac defibrillator infections. Infection limited to the generator pocket may cause pain, redness and swelling that is often accompanied by drainage or fistula formation. In this instance, the generator should be removed and reimplanted at another site as cure is rare with antimicrobial therapy alone. Infection of the leads usually tracks along the wire to include the endocardial surface and may involve the tricuspid valve and pocket. Clinical manifestations vary from mild chronic non-specific symptoms to septic shock with marked localizing signs. Septic embolization to the lungs is common and may cause cough, chest pain and shortness of breath that may be misdiagnosed. Blood culture and trans-oesophageal echocardiography (TOE) are the most important investigations.TOE has a sensitivity of >90%. Lead infection without vegetations may occur and these infections should be treated as for endocarditis. Antimicrobial therapy is an important part of treatment but lead infections are unlikely to cured unless the device is removed. Vancomycin is suitable as initial antimicrobial therapy as this covers both S. aureus and coagulase-negative staphylococci. Flucloxacillin, dicloxacillin or a first-generation cephalosporin are preferred if the organism is sensitive. The addition of low-dose gentamicin may improve bacterial killing. The duration of antimicrobial therapy and timing of replacement of the device have not been determined but 2 weeks treatment before removal and 2-4 weeks treatment after replacement is commonly administered.

Anti-Bacterial Agents↗

[Staphylococcal infection in the newborn: teicoplanin therapy].

Infections caused by Gram-positive bacteria, particularly in neonatal patients, have increased dramatically over the past 10 years. In the present study 19 newborns (7 at term, 12 preterm) with proven staphylococcal infection were treated with teicoplanin, after a previous ineffective antibiotic treatment (amikacin+oxacillin or third-generation cephalosporin). Bacterial eradication and clinical cure were achieved in all neonates. No adverse events related to the drug occurred. No significant change was observed in serial biochemical and hematological tests. Our results suggest that teicoplanin is highly effective and safe in neonatal staphylococcal infections.

Anti-Bacterial Agents↗

Antibodies to Staphylococcus aureus peptidoglycan and lipoteichoic acid in sera from blood donors and patients with staphylococcal infections.

An enzyme-linked immunoassay (ELISA) was used to detect antibodies in human sera to Staphylococcus aureus peptidoglycan (PG) and lipoteichoic acid (LTA). All the sera from the blood donors contained IgG antibodies to both substances. Among the sera from 34 patients with bacteriologically verified, serious S. aureus infections, 71 per cent contained significantly elevated levels of anti-PG antibodies and 76 per cent of anti-LTA antibodies. Among the sera from 38 patients with suspected but not bacteriologically verified staphylococcal infections, 58 per cent contained significantly elevated levels of anti-PG antibodies and 74 per cent of anti-LTA antibodies. The levels of antibodies to PG correlated well with the levels of antibodies to LTA, but the latter occurred over a broader range in the patient sera. Elevated antibody values were, however, also found in some patients with serious, non-staphylococcal infections. The diagnostic value of PG and LTA antibodies has to be further investigated.

Antibodies, Bacterial↗

[The mast cell reaction to a staphylococcal infection occurring against a background of immunodeficiency after cyclophosphane administration].

Materials on the study of the morphofunctional state of mast cells in mouse in experimental staphylococcal infection under the conditions of cyclophosphamide-induced immunodeficiency are presented. As revealed in this study, the infectious process developing in the presence of immunodeficiency is accompanied by the profound and prolonged suppression of the morphofunctional status of mast cells and natural immunity factors at the peak of the disease.

Animals↗

[Contribution of bactericidal rate in the initial treatment of severe staphylococcal infections in children].

Rapid bactericidal effect is essential for therapy of severe Staphylococcus aureus infections in children. The in vitro activity of clinically achievable levels of vancomycin and its combinations with gentamicin, amikacin and rifampin plus amikacin was studied with the time-kill curve method at 2.5, 4, 6, 24 and 48 hours against 20 strains of Staphylococcus aureus isolated from children with severe staphylococcal infection. Vancomycin alone exerted a bactericidal effect at 48 hours. However, with the combinations vancomycin plus gentamicin, vancomycin plus amikacin and vancomycin plus amikacin plus rifampin a bactericidal effect was respectively observed at 24, 6 and 4 hours. Most rapid killing was achieved with vancomycin plus rifampin plus amikacin. Thus this antibiotic combination seems the most appropriate for initial treatment of severe staphylococcal infections in children.

Anti-Bacterial Agents↗

Treatment of severe staphylococcal infections with cefotaxime and fosfomycin in combination.

In a prospective study, 23 severe staphylococcal infections (9 meningitis, 10 bone and joint infections, 3 septicaemia, 1 superinfection of a congenital varicella) were treated with cefotaxime and fosfomycin in combination. There was a synergistic effect of the combination for 14 of the 17 strains tested. Three hours after the end of the infusion, mean CSF concentrations of cefotaxime and fosfomycin were respectively at day 2 3.2 mg/l and 31.4 mg/l, at day 4 2.9 mg/l and 33.9 mg/l. All the patients with meningitis or acute bone and joint infections recovered satisfactorily without relapses. Two superinfections were observed (one systemic candidosis and one septicaemia due to Pseudomonas aeruginosa). Clinical tolerance was good and treatment was discontinued for side effects in only three patients.

Adolescent↗