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It appears to be entirely appropriate for pharmacists to administer vaccinations if restricted to a limited number of vaccines and a well-defined set of recipients. Recommended types of vaccines would be inert vaccines with no contraindications, including flu vaccines, booster shots for diphtheria, tetanus, pertussis, and polio, and HPV vaccines for the prevention of cervical cancer. Recipients targeted for these types of vaccinations would only be adults and adolescents. In addition, pharmacist-administered vaccinations would not be recommended for pregnant women, people with immunodeficiencies, chronic diseases, or cystic fibrosis, people under treatment (anticoagulants) or with known allergies, and haemophiliacs. They would not be recommended either when needed in the context of employment and for traveling abroad. Training is essential to manage the successful implementation of a pharmacist-administered vaccination program (maintaining cold storage, monitoring, space allocation, vaccination administration process, preventive measures, quick recognition and management of anaphylactic chock…).

Summary Background and objectives: Levobupivacaine in combination with sufentanil may be used for labour or postoperative regional analgesia. The risk of bacterial growth within these contained solutions for several hours at room temperature is unknown. We investigated the in vitro antimicrobial effect of levobupivacaine and sufentanil against common micro-organisms encountered during regional anaesthesia. Methods: Standardized suspensions of Staphylococcus aureus, Staphylococcus epidermidis and Escherichia coli were incubated for 1, 3, 6 and 24h at 258C, with saline (as control), sufentanil 0.5 or 0.75 mgmL 21 , levobupivacaine hydrochloride 5.6mgmL 21 and concentrations of 1.4, 2.8 and 5mgmL 21 of levobupivacaine hydrochloride with sufentanil 0.5 mgmL 21 . Colony counts were compared after 24h incubation at 378C. Results: No bacterial growth was observed on any bacterial strain for any solution tested throughout the experiment. Conclusions: Our results suggest that solutions of levobupivacaine combined with sufentanil may be used for 24h at room temperature during regional anaesthesia with no risk of bacterial growth.

In sportswear industry, body odors generated, in particular, in shoes are a big issue that remains unsolved because of the difficulties to destroy odors and bacteria at the same time. The effects of TiO 2 coated textiles for footwear application on bacteria degradation have been assessed in darkness and under UV-A ( λ > 340 nm) irradiation, using Staphylococcus epidermidis as model microorganism of skin bacteria. TiO 2 treated polyester textiles were impregnated by padding process using TiO 2 Degussa P-25 suspensions. The antibacterial effects of these textiles were evaluated using ISO 20743 standard with the transfer method and quantified by the plate count method. This standard has been adapted for photocatalytic tests on bacteria, adding an irradiation step to the ISO 20743 experimental procedure and new equations. In this study, different irradiances, exposure times, and titania contents were compared to find optimal performances on bacteria degradation. A total innocuity on skin bacteria of TiO 2 coated textile was found in the dark. By contrast, under UV-A irradiation, even at very low irradiance (0.13 mW/cm 2 ) of 20 min of UV exposure allows to completely inactivate S. epidermidis cells and avoid their regrowth with the 5.7 g/m 2 TiO 2 -coated polyester surface. After 1 h of UV exposure, whatever the amount of TiO 2 used (1–6 g/m 2 ), all treated textiles were bactericidal.

Resume La PVP-I est un antiseptique a large spectre dont l'activite antimicrobienne a ete largement demontree. Les solutions de PVP-I dermique ou moussante sont bactericides selon les normes AFNOR en 5 minutes. Des etudes in vivo ont montre une efficacite apres 5 min, maximum a 15 min, persistante a une heure. Des essais in vitro ont montre une activite bactericide en 15 secondes, ce qui laisse presager que l'antisepsie de la peau saine pourrait etre effective tres precocement. Pour mettre en evidence cette rapidite d'action in vivo, nous avons donc effectue une cinetique de bactericidie de la PVP-I solution dermique, aux temps 30 sec, 1, 2 et 5 minutes, apres application unique de 15 et 30 secondes, sur le pli du coude de 24 sujets sains. Les prelevements cutanes ont ete effectues par la technique du lavage, dans un liquide neutralisant contenant du thiosulfate. Un abaissement de 1,4 log10 du nombre de colonies bacteriennes par cm2 de peau a pu etre obtenu en 15 secondes sur la flore aerobie et de 1 log10 sur la flore aero-anaerobie du pli du coude. Ces resultats devraient permettre une utilisation de la PVP-I solution dermique pour l'antisepsie rapide de la peau avant des gestes de soins tels que des prelevements sanguins ou des injections medicamenteuses.

Chlorhexidine is an antiseptic widely used to clean the skin. The present work compare the percutaneous diffusion of Hibitane® through hairless rat skin with or without stratum corneum. For the tests carried out on whole skin, the storage is more important than the diffusion; the reverse was observed for stripped skin. The results are discussed according the composition of the commercial solution and the physicochemical characteristics of chlorhexidine digluconate.

The MEGAPOLI experiment took place in July 2009. The aim of this campaign was to study the aging and reactions of aerosol and gas-phase emissions in the city of Paris. Three ground-based measurement sites and several mobile platforms including instrument equipped vehicles and the ATR-42 aircraft were involved. We present here the variations in particle- and gas-phase species over the city of Paris using a combination of high-time resolution measurements aboard the ATR-42 aircraft. Particle chemical composition was measured using a compact time-of-flight aerosol mass spectrometer (C-ToF-AMS) giving detailed information of the non-refractory submicron aerosol species. The mass concentration of BC, measured by a particle absorption soot photometer (PSAP), was used as a marker to identify the urban pollution plume boundaries. Aerosol mass concentrations and composition were affected by air-mass history, with air masses that spent longest time over land having highest fractions of organic aerosol and higher total mass concentrations. The Paris plume is mainly composed of organic aerosol (OA), black carbon and nitrate aerosol, as well as high concentrations of anthropogenic gas-phase species such as toluene, benzene, and NOx. Using BC and CO as tracers for air-mass dilution, we observe the ratio of ΔOA / ΔBC and ΔOA / ΔCO increase with increasing photochemical age (−log(NOx / NOy). Plotting the equivalent ratios for the Positive Matrix Factorization (PMF) resolved species (LV-OOA, SV-OOA, and HOA) illustrate that the increase in OA is a result of secondary organic aerosol (SOA). Within Paris the changes in the ΔOA / ΔCO are similar to those observed during other studies in Mexico city, Mexico and in New England, USA. Using the measured VOCs species together with recent organic aerosol formation yields we predicted ~ 50% of the measured organics. These airborne measurements during the MEGAPOLI experiment show that urban emissions contribute to the formation of OA, and have an impact on aerosol composition on a regional scale. They provide a quantitative measure of this impact in terms of urban plume composition and evolution relative to background aerosol composition.

Twenty-five strains of staphylococci isolated from goat milk and cheese were identified as belonging to the Staphylococcus xylosus/equorum group using the ID 32 Staph system (bioMerieux, Marcy-L'Etoile, France). This system, however, was not able to discriminate between these two species for 19 of the strains tested. Ribotyping was performed on these 25 strains, as well as on three reference strains of each of these two species. Hybridization membranes were scanned and analyzed using the Taxotron® software package (Taxolab, Institut Pasteur, Paris, France). A dendrogram representation showed that ribotypes were distributed in two clear-cut clusters corresponding to S. equorum (21 strains) and S. xylosus (four strains).

The Wellcolex Colour Salmonella Test (WCST) is a rapid latex agglutination test designed to detect Salmonella antigens in stool enrichment broth cultures or from plate cultures. As the reagents are mixtures of suspensions of red, blue, and green latex particles, serogroup identification can be achieved in a single test. The results of the WCST using overnight selenite broth cultures were compared with standard culture results. A total of 702 stool samples were evaluated in this study. Thirty-nine Salmonella strains (5.5%) were isolated while 50 positive results were obtained with the WCST reagents. In 14 other cases, the latex test could not be interpreted because of doubtful agglutination. Excluding these cases, the specificity of the WCST was 98% and the percentage agreement between the WCST and teh culture rsults was 98%. The predictive value of a positive test was 78%. Of particular note, the predictive value for negative results was 100% showing that the WCST can be used to effectively predict those samples which do not need further workup. The color change (red, blue, green) is easy to observe and can identify 6 serogroups (A–E and G). We conclude that the WCST provides a simple and rapid procedure for detection and presumptive serogroup identification of Salmonella spp. in enrichment broths.

Resume Les bacilles a Gram negatif oxydase-positive du genre Flavobacterium sont habituellement decrits comme multiresistants aux antibiotiques posant le probleme de leur prevalence future comme opportunistes, en particulier pour F. meningosepticum, germe le plus souvent isole. Nous avons examine le comportement de 121 souches de Flavobacterium vis-a-vis de 39 antibiotiques dont 14 β-lactamines par la methode de diffusion : 112 souches sont multiresistantes a l'exception de quelques antibiotiques : ureidopenicillines, minocycline et rifampicine. Une synergie entre l'amoxicilline ou la ticarcilline et l'acide clavulanique est toujours observee a l'exception des souches de F. odoratum. En outre, seule l'espece F. meningosepticum presente une synergie entre cet inhibiteur et les cephalosporines telles la ceftazidime, le cefotaxime. L'analyse exploratoire des donnees par la methode graphique (MacSpin) et mathematique (ACP) de 14 β-lactamines confirme l'individualisation prealable de phenotypes de resistance par espece (F. meningosepticum, F. indologenes, F. breve, F. odoratum, F. multivorum et F. spiritivorum) permettant une identification plus rapide de ces germes inhabituels.

A group of 17 strains of coagulase negative staphylococci isolated from slaughtered lambs, and which could not be identified with the conventional methods, exhibited high levels of DNA homology (92%) with the S. caseolyticus reference strain. The isolates described in this study provide a more extensive comprehension of S. caseolyticus. The original description of this species was based on only two strains isolated from milk. To our knowledge, S. caseolyticus had never been previously associated with animal microflora.

Adhesion of bacteria to intraocular lenses is an important step in the pathogenesis of postoperative endophthalmitis. It can be described as a two-phase process including an initial, instantaneous, and reversible phase followed by a time-dependant and irreversible molecular and cellular phase. The binding of bacteria is affected by many factors including environmental factors such as medium composition, presence of proteins and flow conditions, the bacterial cell surface characteristics, and the material's surface properties. This article reviews all these factors affecting the adhesion of bacteria to intraocular lenses. A better understanding of these mechanisms would make it possible to reduce the bacterial adhesion process and thus could help decrease the incidence of postoperative endophthalmitis.

Cataract surgery is a usually successful procedure that restores vision by replacing the natural lens with an intraocular lens (IOL). Acute postoperative endophthalmitis is still one of the most serious complications of cataract surgery. Its incidence has been reported to be between 0.04% and 0.32%. Precisely why bacteria induce endophthalmitis is not entirely understood. Indeed the risk of its development may be influenced by several factors. Among them, bacterial adhesion to the IOL has been recently emphasized in the ophthalmology literature. Indeed, the ability of an organism to adhere to the IOL surface is believed to be associated with a risk of infection at the implantation site. Several studies have demonstrated that bacterial adhesion is influenced by IOL materials. Ever since, numerous studies have investigated the interactions between bacteria and different types of IOLs to determine which biomaterial would be most permissive to bacterial adherence. This article reviews all the epidemiological and experimental data relating to the study of the relationship between bacterial adhesion, IOL material, and risk of developing postoperative endophthalmitis. Even if discrepancies between these studies exist, mainly stemming from the use of different experimental conditions and protocols, it seems that bacterial adhesion is strongly influenced by IOL material. Epidemiological studies suggest that the implantation of silicone IOLs might be associated with increased rates of endophthalmitis. Experimental studies reach similar conclusions showing that hydrophobic IOLs such as silicone or acrylic hydrophobic IOLs are more permissive to bacterial adhesion and growth than hydrophilic IOLs such as acrylic hydrophilic IOLs. Among the interactions that govern bacterial attachment to the IOLs, it seems that hydrophilic-hydrophobic interactions have the greatest influence. Nevertheless, since bacterial adhesion is a complicated process affected by many factors, the conclusions drawn by these results have to be interpreted with care. Further investigations are still needed to understand the connections between IOL material and endophthalmitis.

Resume Un cas d'infection simultanee par deux souches distinctes de Staphylococcus epidermidis chez une patiente immunodeprimee est rapporte. Les clones bacteriens ont ete caracterises a l'aide des marqueurs epidemiologiques conventionnels, mais aussi moleculaires comme l'etude des profils plasmidiques et des profils de restriction de l'ADN total apres utilisation d'une endonuclease.

Efflux pumps located in the bacterial membranes are responsible for low level resistance to antibiotics, considered not to be relevant in the clinic and thus often neglected. However, these pumps contribute to the emergence of high level antibiotic resistance mechanisms, which are responsible for severe complications during the treatment of infectious diseases. Therefore it is necessary to take into account these pumps while developing novel antibacterial agents. Among these new research strategies, the development of efflux pump inhibitors seems to be an attractive approach to restore the activity of some "classical" antibiotics and to limit the emergence of multiresistant strains associated with hospital-acquired infections. In this review, we focalise on Staphylococcus aureus efflux pumps and their potential inhibitors.

Resume En 1899 et 1900, Tissier met en evidence les premiers isolats de souches provenant de matieres fecales de nourrissons, qui sont decrits a partir de 1924, comme appartenant a un genre nouveau, le genre Bifidobacterium. Le genre comprend aujourd'hui 26 especes, dont 10 sont considerees comme specifiquement d'origine humaine, 14 d'origine animale, et 2 provenant de l'environnement hydrique. Cette revue presente l'evolution du genre depuis son origine, et les methodologies qui sont a la base de la creation de ces differentes especes. Les donnees sur le genre Bifidobacterium sont critiquees en fonction de l'apport des techniques actuelles de classification, phenotypique et genomique (analyse numerique, hybridation ADN-ADN) et des systemes d'identification fondes sur le calcul des probabilites, developpes pour de nombreux autres groupes bacteriens.

Resume Une bacterie presentant l'ensemble des caracteristiques de Rhodococcus equi a ete isolee d'une hemoculture et d'un catheter chez un homme de 58 ans atteint d'une leucemie aigue myeloblastique. L'observation clinique est rapportee et les caracteristiques physiologiques et biochimiques ainsi que l'etude de la composition des acides gras cellulaires de cette souche sont decrites dans la presente note. Une revue de la litterature est egalement presentee.

In the last few years, regulations for biomolecule production, and especially for extraction and purification of animal molecules such as collagen, have been reinforced to ensure the sanitary safety of the materials. To be authorized to market biomaterials based on collagen, manufacturers now have to prove that at least one step of their process is described in guidelines to inactivate prion, viruses, and bacteria. The present study focuses on the inactivation step performed during the extraction and purification of porcine type I atelocollagen. We chose to determine the reduction factor of a 1 M NaOH step on porcine parvovirus and four bacterial strains inactivation. During the extraction step, we deliberately inoculated the collagen suspension with the different microorganisms tested. Then, 1 M NaOH was added to the suspension for 1 hour at 20 degrees C. We demonstrated that this treatment totally inactivated S. aureus, P. aeruginosa, C. albicans and A. niger which are bacterial strains responsible of severe human pathology. The reduction factors reached more than 4 logs for B. cereus spores and 4 logs for the porcine parvovirus. are encouraging as those two microorganisms are known to be very resistant to inactivation.

Bacterial adhesion to intraocular lenses (IOLs) during their implantation is a major etiological factor of postoperative endophthalmitis. Polypropylene was the first biomaterial that allowed this relation of cause and effect between bacterial adhesion and endophthalmitis to be proven. Adhesion to IOLs has been investigated in several in vitro studies, with contradictory results reported, due to variations in experimental conditions: the incubation times and the methods all varied. Adhesion is affected by the type of IOL, the bacterial strains, and the surrounding medium. Since this medium is very difficult to model because of its complexity, in vivo studies seemed essential. We have recently determined in vivo progression in the amount of adhering Staphylococcus epidermidis to five types of IOLs. There have been few epidemiological studies published to determine the relationship between endophthalmitis and the IOL type. However, the perfect biomaterial that could prevent postoperative endophthalmitis does not yet exist. Globally, hydrophilic materials and hydrophobic acrylic seem to be less sticky than silicone or PMMA, but this remains to be proven clinically.

Postoperative endophthalmitis is still one of the most fearsome complications of cataract surgery. The present review's aims are to study the etiology and pathogenesis of endophthalmitis and the criteria for antibiotic prophylaxis. Endophthalmitis prevalence is 0.07%-0.32% in cataract surgery. The clinical presentation needs to be perfectly known, even if none of the signs is pathognomonic. Bacteria predominantly cultured in postoperative endophthalmitis are gram-positive, especially Staphylococcus epidermidis. Most of the bacteria come from the patient. Bacterial adhesion to intraocular lenses (IOLs) takes place during their implantation, a prominent etiological factor. Polypropylene was the first biomaterial that proved this relation of cause and effect between bacterial adhesion and endophthalmitis. The benefit of antibiotic prophylaxis during cataract surgery has yet not been proven, since the low prevalence of endophthalmitis makes controlled studies with a large cohort difficult. The criteria and the four possible administration approaches (topical, subconjunctival, irrigation liquid, systemic) are analyzed.

Bacterial adhesion to intraocular lenses (IOLs) takes place during their implantation. This is a prominent etiological factor of postoperative endophthalmitis. Following adhesion, secretion of an extracellular matrix (called slime for Staphylococcus epidermidis) and formation of multiple layers of microcolonies lead to the colonization of the biomaterial surface. Scanning electron microscopy photographs illustrate the different steps of biofilm formation. The different adhesins expressed by S. epidermidis involved in the adhesion process are described. The biofilm is not only an adhesive medium; it also affects virulence. Last, notions on biofilm physiology are discussed in an attempt to explain the dynamic equilibrium of this system. In 2004, the perfect biomaterial able to prevent postoperative endophthalmitis does not yet exist. Moreover, there is no effective tool, at the present time, to fight against mature biofilms. Therefore, preventing biofilm formation remains capital, which requires perfect knowledge of all stages of formation and the factors involved.

API Candida was evaluated in comparison with the ID 32C system for the identification of 619 yeast isolates. The sensitivity of API Candida for the identification of the 15 species it claims to identify with and without additional tests was 97.4% (593 of 609) and 75.2% (458 of 609), respectively. The API Candida system is easy to use and rapid (result in 18 to 24 h).

Postoperative endophthalmitis following intraocular lens (IOL) implantation is still one of the most feared complications of cataract surgery. Bacterial adhesion to IOLs during their insertion is a prominent etiological factor. Polypropylene was the first biomaterial that allowed this relation of cause and effect to be proven. Following adhesion, bacteria replicate, congregate and form multiple layers of microcolonies which actually represent the basic structural unit of the biofilm. The bacteria are embedded in a slime layer. Personal photographs illustrate the different steps of biofilm formation. This slime matrix is not only an adhesive medium; it also affects virulence. Adhesion to IOLs has been studied by several in vitro studies and discrepancies can be found between them which are due to variations of experimental conditions. The strains, the incubation times and the methods all varied. Adhesion is affected by the nature of the IOLs, the isolates and the surrounding medium. Since this medium is very difficult to model because of its complexity, in vivo studies seemed essential. We have recently determined in vivo evolution of the amount of attached bacteria to five types of IOLs. Crystalline lenses from 90 domestic pigs were removed aseptically and replaced with previously infected IOLs. There have been few epidemiological studies published to determine the relationship between endophthalmitis and the IOL type. However, the perfect biomaterial that could prevent postoperative endophthalmitis does not yet exist. Globally, hydrophilic materials and hydrophobic acrylic seem to be less sticky than silicone or PMMA, but this remains to be proven clinically.

CPS ID2 medium (bioMérieux) enables the presumptive identification of Escherichia coli and enterococci as well as the detection of indologenous or nonindologenous Proteeae and bacteria belonging to the Klebsiella, Enterobacter, and Serratia group with a specificity ranging from 98 to 100%. When the cultures were polymicrobial, the sensitivity varied from 70 to 97%, and the sensitivity varied from 97 to 100% when they were monomicrobial.

Advances in biotechnologies and miniaturization over the past few decades have led to an explosion in the number of rapid tests in in vitro diagnosis and explain their ever-growing presence in international markets. Although based on classic immunological and biochemical methods, these tests deliver many more benefits for laboratories. They are rapid and simple to perform, provide results rapidly and can be performed in remote laboratories, making them invaluable for both routine and emergency analyses. As beneficial as they may be, however, they still have a long way to go before they find widespread use in infectious disease control. Currently, rapid testing methods are used only to screen for urinary tract infections, monitor certain infectious parameters and make more accurate diagnosis of a still small number of species of bacteria. This narrow catalogue of available tests is yet another disadvantage in a list of drawbacks typically associated with rapid tests. A good portion of complaints centers on the fact that they are still too recent to have sufficient information on their levels of sensitivity and reliability. Furthermore, regulations, in particularly concerning accountability, remain to be established. Nevertheless, constant innovation coupled with the experience of laboratory professionals in the field should open the way for rapid testing to earn its place in the line-up of medical laboratory tests.

Over a 16-month period from September 1997 to December 1998, a prospective study was made of an on-site treatment of medical wastes in a 10-bed intensive care unit. First, the wastes were ground and then, a high concentration of ozone in air was repeatedly injected into the ground wastes. The study analysed the practical application of the system and its microbiological efficiency. Inactivation experiments were made with reference strains of Staphylococcus aureus , Enterococcus hirae , Pseudomonas aeruginosa , Escherichia coli , Mycobacterium smegmatis , Bacillus subtilis var niger , Bacillus stearothermophilus , Candida albicans and Aspergillus niger . Two thousand eight hundred treatment cycles, i.e. 84000 grindings and 140000 ozone injections gave a treatment capacity of 50kg of waste per day with a good staff acceptability. All kinds of medical devices used in an intensive care unit were treated. In untreated ground wastes, the median bacterial load was 105.86 (range 10 2.35 –10 8.05 ) cfu/g. After ozone treatment, bacteria and fungi were reduced by a factor of 10 5 . Aero-contamination of the ward was unchanged. Computer control allowed all events to be tracked. On-site medical waste treatment appears to be an efficient alternative to the usual centralized collection and treatment.

In any health care institution, the most important quality indicator for antisepsis is the existence of systems of reference of use, secondly the knowledge of those systems and then the compliance by the different classes of professionals. These refrentials must be regularly evaluated and updated according to general consensus.

Summary Sequencing and comparison of 16S rRNA genes of Staphylococcus haemolyticus, S. hominis and S. carnosus allowed the development of an oligonucleotide probe that is complementary to a region of S. haemolyticus 16S rRNA and allows the differentiation of this species from the other two. After hybridization with 27 staphylococcal reference strains and 63 clinical strains, we found this probe useful as an alternative to the ID 32 Staph gallery (bioMerieux) in the case of an identification problem between S. haemolyticus and S. hominis or S. warneri . The oligonucleotide probe was used for the characterization of two misidentified strains.

In accordance with Recommendation 30b of the International Code of Nomenclature of Bacteria, minimal standards are proposed for the genus Staphylococcus and the description of newly recognized species in this genus. Assignment of a strain to the genus Staphylococcus requires that it is a Gram-positive coccus that forms clusters, produces catalase, has an appropriate cell wall structure (including peptidoglycan type and teichoic acid presence) and G + C content of DNA in a range of 30-40 mol%. The recommended minimal standards for describing a new Staphylococcus species are based on the results of phenotypic and genomic studies of at least five independently isolated strains. They include colony morphology and the results of the following conventional tests: pigment production, growth requirements, fermentative and oxidative activity on carbohydrates, novobiocin susceptibility, enzymic activities (nitrate reductase, alkaline phosphatase, arginine dihydrolase, ornithine decarboxylase, urease, cytochrome oxidase, staphylocoagulase in rabbit plasma, heat-stable nuclease, amidases, oxidases, clumping factor, and haemolytic activity on sheep or bovine blood agar). DNA-DNA hybridization experiments may distinguish species when the difference between the binding in the homologous reaction and the binding in the heterologous reaction expressed as a percentage is less than 70%. In addition, rRNA signature sequence criteria, ribotyping characterization of the nomenclature type strain and other strains of the species, and reference strains of other species is recommended to describe the strains of the new species with sets of genetic attributes and reveal possible grouping errors. This proposal has been endorsed by the members of the Subcommittee on the taxonomy of staphylococci and streptococci of the international Committee on Systematic Bacteriology.

Residual chlorhexidine concentrations were measured after application of a single dose on the skin of 22 healthy volunteers. Dosage by high-pressure liquid chromatography in the skin cleansers revealed that the residual concentrations were higher than chlorhexidine MICs for most organisms of the resident skin flora and some responsible for hand-borne infections, even 24 h after application.

In a multicenter study, 407 strains of coryneform bacteria were tested with the updated and extended API (RAPID) Coryne system with database 2.0 (bioMérieux, La-Balme-les-Grottes, France) in order to evaluate the system's capability of identifying these bacteria. The design of the system was exactly the same as for the previous API (RAPID) Coryne strip with database 1.0, i.e., the 20 biochemical reactions covered were identical, but database 2.0 included both more taxa and additional differential tests. Three hundred ninety strains tested belonged to the 49 taxa covered by database 2.0, and 17 strains belonged to taxa not covered. Overall, the system correctly identified 90.5% of the strains belonging to taxa included, with additional tests needed for correct identification for 55.1% of all strains tested. Only 5.6% of all strains were not identified, and 3.8% were misidentified. Identification problems were observed in particular for Corynebacterium coyleae, Propionibacterium acnes, and Aureobacterium spp. The numerical profiles and corresponding identification results for the taxa not covered by the new database 2.0 were also given. In comparison to the results from published previous evaluations of the API (RAPID) Coryne database 1.0, more additional tests had to be performed with version 2.0 in order to completely identify the strains. This was the result of current changes in taxonomy and to provide for organisms described since the appearance of version 1.0. We conclude that the new API (RAPID) Coryne system 2.0 is a useful tool for identifying the diverse group of coryneform bacteria encountered in the routine clinical laboratory.

An upsurge in pertussis infections, despite mandatory vaccination in France since 1966, has occurred again in developed countries due to progressive loss of vaccinal immunity and wider circulation of the causal bacterium, Bordetella pertussis. Unfortunately, the classical culture method is insufficiently sensitive and serology can only confirm diagnosis retrospectively. New techniques are needed for rapid diagnosis, and subsequent treatment and preventive measures. One new method, gene amplification using polymerase chain reaction (PCR), has been particularly useful in detecting Bordetella pertussis. PCR is highly specific and more sensitive than culture. It is thus quite useful in case of atypical clinical presentations and in previously treated or vaccinated patients. Less restrictions on sample transportation and preservation make PCR a technique which general practitioners can use for rapid easy diagnosis of pertussis.

Staphylococcus aureus is one of the prominent causative agents of ventilator-associated pneumonia (VAP). Gram staining of bronchoalveolar lavage (BAL) fluid is not always reliable. A nonisotopid probe (Accuprobe) has been developed by Gen-Probe for the specific identification of S. aureus isolated from cultures. This study was undertaken to assess the reliability of this probe for the early diagnosis of S. aureus VAP.A prospective study in 120 consecutive patients.Department of intensive care medicine at a university hospital.120 ventilated patients (70 males and 50 females; mean age 52 +/- 12 years; mean simplified acute physiologic score = 13 +/- 4) were studied.164 bronchoalveolar lavages were performed (none of the patients received prior antibiotic therapy).S. aureus was identified 29 times at significant concentrations (or = 10(4) cfu/ml) and 7 times at10(4) cfu/ml. The sensitivity and specificity of the Accuprobe system were 100 and 96%, respectively. We found agreement between quantitative cultures and probes in 96.3% of cases.We conclude that this probe provides a rapid (or = 7 h) and accurate diagnosis of S. aureus pulmonary infection.

The mecA gene which confers the oxacillin resistance has been searched by PCR in 290 (124 positives, 166 negatives) coagulase-negative staphylococci (CNS) belonging to twelve species. The results were compared with the oxacillin MIC values obtained by agar dilution (4% NaCl) or by the ATB STAPH method (Api-bioMérieux; 0%, 2%, 5% NaCl) and growth inhibitory diameters obtained by agar diffusion with an oxacillin disk placed at 30 degrees C without NaCl, or at 35 degrees C in presence of 2% or 5% NaCl. Sensitivity of oxacillin resistance detection depends upon the salt concentration and the method used. The optimum concentration is 2%. With this concentration, the Api ATB test appears as the more performant (sensitivity: 89.8%). Search for the mecA gene by PCR represents a very interesting method that detects 96.9% of the oxacillin-resistant CNS strains.

A polymerase chain reaction (PCR) assay was developed for the detection in clinical samples of mycobacteria belonging to the Mycobacterium tuberculosis complex. PCR products were detected with a simple and rapid colorimetric method. With this method, 50 fg of M. tuberculosis DNA were detectable with the repetitive DNA-sequence-derived primers, corresponding to 10 genome equivalents. Detection of M. tuberculosis in 258 clinical samples by PCR was compared with detection by culture. PCR was positive for 56 of 57 culture-positive and Ziehl—Neelsen-staining-positive (ZN) samples, 11 of 18 culture-positive and ZN-negative samples. The presence of gro EL DNA sequences was also investigated by PCR for all the specimens with the same revelation protocol. Three of the eight false-negative samples with the repetitive element-derived primers were found to contain gro EL DNA sequences specific for the Mycobacterium genus. Among the 183 culture-negative samples, 30 were positive by PCR. When clinical data were known, the diagnosis of tuberculosis was established for the patients from whom those samples had been obtained. The results show that the rapid and simplified PCR assay described here is slightly more sensitive than culture and can be used in routine clinical practice.

The Polymerase Chain Reaction (PCR) was used to detect the qacA gene which encodes antiseptic resistance in 186 clinical isolates of Staphylococcus aureus. The results were compared with those obtained by MIC determination of 4 antiseptics (benzalkonium chloride, hexamidine, chlorhexidine, acriflavine) and for ethidium bromide. The qacA gene was not detected among the 32 susceptible S. aureus strains, but was found in the 70 (85%) of the 82 S. aureus strains resistant to all 5 antiseptics. The gene was also detected in 70 (45%) of the 154 remaining strains that were resistant to at least one antiseptic.

Conventional methods usually fail to identify Klebsiella planticola and Klebsiella terrigena, which represent up to 19% of clinical Klebsiella isolates. By combining four carbon substrate assimilation tests and two conventional tests, the method identified these species with a specificity and a sensitivity of 100%. Overall, sensitivity for Klebsiella identification was 94.7%.

Vibrio metschnikovii is largely distributed in the aquatic environment; human infections are rarely observed. A fatal case of septicemia in a patient with liver cirrhosis, renal insufficiency, and diabetes is described. A second case in a 82-year-old woman with septicemia, respiratory problems, and infected leg lesions is reported; she was successfully treated.

The effect of six representative cell wall-active antibiotics upon the membrane proteins of Staphylococcus aureus CIP 65-25 was studied. The profiles of whole membrane proteins and of penicillin-binding proteins of strain CIP 65-25 grown on an antibiotic-free medium were analysed and compared with those of nine other S. aureus strains. Both profiles were found to be very similar for all strains examined. Cell wall-active antibiotics brought about definite changes dealing with a few membrane proteins of strain CIP 65-25. Some polypeptide bands of the control profiles decreased or vanished after exposure to antibiotics whereas others rose only in antibiotic-treated bacteria. None of these modified polypeptides was a penicillin-binding protein. Their possible involvement in the antibacterial effects of antibiotics or in the determinism of bacterial resistance to antibiotics is discussed.

A non-isotopic nucleic probe (Accuprobe) has been presented recently by Gen-Probe for the direct specific 1-h identification of Staphylococcus aureus isolated from culture. 50 S aureus strains previously characterized by conventional methods as well as 26 atypical strains (absence of coagulase, thermonuclease and fibrinogen affinity factor) were tested. Moreover, the Accuprobe system was evaluated using 29 other staphylococcal type strains representing all the species described to date. Only the strains that belonged to the S aureus species, including the 26 atypical strains, were detected by Accuprobe, which proved to be a rapid specific mean of identifying S aureus strains, particularly those that are not readily identified by conventional methods.

A number of studies have underlined the interest of the polymerase chain reaction (PCR) in the detection of Mycobacterium tuberculosis in clinical samples. Among the different parameters to be carefully studied the choice of target gene and primers is essential. The amplification of nucleotidic sequences localised on three different target genes (groEL, IS6110, Pab) was examined in 196 clinical samples from patients with suspected tuberculosis or receiving antituberculous therapy. The results obtained after hybridization with non-radioactive labelled probes were compared with the culture data. None of the primer sets studied showed a satisfactory sensitivity (79% to 84%) suitable for it to be used alone. The false-negative specimens with the PCR tests usually corresponded to those that contained few mycobacteria. With the methods described in this study, the use of two or three primer sets located on different target genes allowed to improve the positivity rate compared to the culture and sensitivity of the test (90-98%), particularly for paucibacillary samples. On the other hand, the interpretation was easier when concordant results were obtained.