High specificity of line-immunoassay based algorithms for recent HIV-1 infection independent of viral subtype and stage of disease.

BACKGROUND: Serologic testing algorithms for recent HIV seroconversion (STARHS) provide important information for HIV surveillance. We have shown that a patient's antibody reaction in a confirmatory line immunoassay (INNO-LIA HIV I/II Score, Innogenetics) provides information on the duration of infection. Here, we sought to further investigate the diagnostic specificity of various Inno-Lia algorithms and to identify factors affecting it. METHODS: Plasma samples of 714 selected patients of the Swiss HIV Cohort Study infected for longer than 12 months and representing all viral clades and stages of chronic HIV-1 infection were tested blindly by Inno-Lia and classified as either incident (up to 12 m) or older infection by 24 different algorithms. Of the total, 524 patients received HAART, 308 had HIV-1 RNA below 50 copies/mL, and 620 were infected by a HIV-1 non-B clade. Using logistic regression analysis we evaluated factors that might affect the specificity of these algorithms. RESULTS: HIV-1 RNA < 50 copies/mL was associated with significantly lower reactivity to all five HIV-1 antigens of the Inno-Lia and impaired specificity of most algorithms. Among 412 patients either untreated or with HIV-1 RNA ≥ 50 copies/mL despite HAART, the median specificity of the algorithms was 96.5% (range 92.0-100%). The only factor that significantly promoted false-incident results in this group was age, with false-incident results increasing by a few percent per additional year. HIV-1 clade, HIV-1 RNA, CD4 percentage, sex, disease stage, and testing modalities exhibited no significance. Results were similar among 190 untreated patients. CONCLUSIONS: The specificity of most Inno-Lia algorithms was high and not affected by HIV-1 variability, advanced disease and other factors promoting false-recent results in other STARHS. Specificity should be good in any group of untreated HIV-1 patients.

Impact of standardised review of intravenous antibiotic therapy 72 hours after prescription in two internal medicine wards.

Post-prescription review of hospital antibiotic therapy may contribute to more appropriate use. We estimated the impact of a standardised review of intravenous antibiotic therapy three days after prescription in two internal medicine wards of a university hospital. In one ward, we assessed the charts of patients under intravenous antibiotic therapy using a standardised review process and provided feedback to the prescriber. There was no intervention in the other ward. After six months we crossed the allocation between the two wards. In all, 204 courses of antibiotic therapy were included in the intervention periods and 226 in the control periods. Post-prescription review led to proposals for modification in 46% of antibiotic courses. Time to treatment modification was 22% shorter in the intervention periods compared with the control periods (3.9+/-5.2 days vs 5.0+/-6.0 days, P=0.007). Patients included in the intervention group had lower antibiotic consumption than patients in the control group, but the intervention had no significant impact on the overall antibiotic consumption of the two wards.

Usefulness of double locus sequence typing (DLST) for regional and international epidemiological surveillance of methicilin-resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of nosocomial infections worldwide. To differentiate reliably among S. aureus isolates, we recently developed double locus sequence typing (DLST) based on the analysis of partial sequences of clfB and spa genes. In the present study, we evaluated the usefulness of DLST for epidemiological investigations of MRSA by routinely typing 1242 strains isolated in Western Switzerland. Additionally, particular local and international collections were typed by pulsed field gel electrophoresis (PFGE) and DLST to check the compatibility of DLST with the results obtained by PFGE, and for international comparisons. Using DLST, we identified the major MRSA clones of Western Switzerland, and demonstrated the close relationship between local and international clones. The congruence of 88% between the major PFGE and DLST clones indicated that our results obtained by DLST were compatible with earlier results obtained by PFGE. DLST could thus easily be incorporated in a routine surveillance procedure. In addition, the unambiguous definition of DLST types makes this method more suitable than PFGE for long-term epidemiological surveillance. Finally, the comparison of the results obtained by DLST, multilocus sequence typing, PFGE, Staphylococcal cassette chromosome mec typing and the detection of Panton-Valentine leukocidin genes indicated that no typing scheme should be used on its own. It is only the combination of data from different methods that gives the best chance of describing precisely the epidemiology and phylogeny of MRSA.

Application and evaluation of double digest selective label (DDSL) typing technique for Pseudomonas aeruginosa hospital isolates.

This study describes the application and evaluation of a recently developed fast bacterial typing technique (double digest selective label - DDSL) for hospital isolates of Pseudomonas aeruginosa. The protocol was based on a simultaneous double digestion/labelling reaction which was performed in a single reaction tube. After agarose gel separation selectively tagged restriction fragments were transferred using deonised water to a nylon membrane and visualized by a colour reaction. Starting from overnight culture, turn around time using this technique was only 8 h. The DDSL typing technique was applied for 77 hospital isolates. Among them 63 isolates were also typed by PFGE and the typing results were compared with those of DDSL. In conclusion, both techniques discriminated bacterial isolates into the same major clusters. DDSL proved to be as discriminatory as PFGE but much faster and easier to set up in a standard microbiological laboratory.

Changing molecular epidemiology of methicillin-resistant Staphylococcus aureus in a small geographic area over an eight-year period.

The epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) at an international level shows that most MRSA strains belong to a few pandemic clones. At the local level, a predominance of one or two clones was generally reported. However, the situation is evolving and new clones are emerging worldwide, some of them with specific biological characteristics, such as the presence of Panton-Valentine leucocidin (PVL). Understanding these changes at the local and international levels is of great importance. Our objective was to analyze the evolution of MRSA epidemiology at multiple sites on a local level (Western Switzerland) over a period of 8 years. Data were based on MRSA reports from seven sentinel laboratories and infection control programs covering different areas. Pulsed-field gel electrophoresis was used to type MRSA isolates. From 1997 to 2004, a total of 2,256 patients with MRSA were reported. Results showed the presence of four predominant clones (accounting for 86% of patients), which could be related to known international clones (Berlin, New York/Japan, Southern Germany, and Iberian clones). Within the small geographic region, the 8-year follow-up period in the different areas showed spacio-temporal differences in the relative proportions of the four clones. Other international MRSA clones, as well as clones showing genetic characteristics identical to those of community-acquired MRSA (SCCmec type IV and the presence of PVL genes), were also identified but presumably did not disseminate. Despite the worldwide predominance of a few MRSA clones, our data showed that at a local level, the epidemiology of MRSA might be different from one hospital to another. Moreover, MRSA clones were replaced by other emerging clones, suggesting a rapid change.

Blood samples drawn for culture as a surrogate marker for case-mix adjustment of hospital antibiotic use.

Hospital antibiotic consumption is generally adjusted to occupancy. This study hypothesised that the number of blood culture samples could be a surrogate marker for case-mix adjustment. Antibiotic consumption was compared over 16 consecutive trimesters in one medical ward in terms of patient-days or blood culture samples. Compared with patient-days, measurement adjusted to blood culture samples detected three trimesters with an unusually high consumption, and one trimester with consumption falsely classified as high because of a high incidence of infections. Blood culture numbers enabled easy and accurate identification of periods with a drift in antibiotic consumption ina medical ward.

Double-locus sequence typing using clfB and spa, a fast and simple method for epidemiological typing of methicillin-resistant Staphylococcus aureus.

Sequence-based epidemiological typing of methicillin-resistant Staphylococcus aureus (MRSA) has recently been promoted because it results in unambiguous data sets that can be organized in local and global databases. The replacement of previous typing methods, such as the highly discriminatory pulsed-field gel electrophoresis (PFGE), has been attempted with various markers and typing schemes, including spa typing and multilocus sequence typing. However, despite a number of advantages, none of these methods showed convincing evidence for performance in epidemiological typing comparable to that of PFGE. By using three sets of 48 MRSA strains comprising isolates that were (i) genetically highly diverse, (ii) genetically related, and (iii) obtained from long-term carriers, we analyzed the performance of the four highly polymorphic S. aureus markers: clfA, clfB, fnbA, and spa. Typeability, discriminatory power, in vivo stability, and evolution of these markers were compared to those of PFGE. Clearly, none of the markers alone could match the discriminatory power of PFGE (63 genotypes; index of discrimination of 0.96). Instead, this could be achieved by combining markers in pairs. We showed that by using only 3' partial sequences of approximately 500 bp, the majority of each marker's discriminatory power was displayed, and using the partial sequences, the best performance was obtained with the combination of clfB and spa (57 genotypes; index of discrimination of 0.94). Genetic changes were not observed for any of the sequence markers over a period of 3 years and in the case of partial sequences for a period of more than 4 years. This is in contrast to PFGE where changes occurred after several months. The genetic differences found between isolate pairs of long-term carriers and among highly related isolates indicated clonal evolution. A typing scheme based on 500-bp 3' partial sequences of clfB and spa is proposed.

Molecular Epidemiology of Pseudomonas aeruginosa in the Intensive Care Units - A Review.

Pseudomonas aeruginosa is one of the leading nosocomial pathogens in intensive care units (ICU). This opportunist pathogen is commonly recovered from moist environments, and is also found colonizing 2.6 to 24% of hospitalized patients. We reviewed the recent literature that used highly discriminatory typing methods to precisely identify the reservoirs and modes of transmission of this microorganism in the ICU setting. In most ICUs, the endogenous flora was suspected to be the main source of infection compared to exogenous sources (other patients, the contaminated environment such as sinks or taps). However, the percentage of endogenous versus exogenous sources might vary considerably from one setting to another. Reasons for this include the compliance of health care workers to infection control measures, the contamination of the environment, and probably also the biology of the pathogen (intrinsic fitness factors). As P. aerugi-nosa is ubiquitous in the environment and colonizes up to 15% of hospitalized patients, eradication of the reservoir is difficult, if not impossible. Therefore, efforts should primarily focus on reinforcement of infection control measures to limit its transmission.

Evaluation of three chromogenic media (MRSA-ID, MRSA-Select and CHROMagar MRSA) and ORSAB for surveillance cultures of methicillin-resistant Staphylococcus aureus.

Screening specimens were homogenised in saline 0.9% w/v before either direct inoculation or following enrichment in broth on three chromogenic media (MRSA-ID, CHROMagar MRSA and MRSA Select) and ORSAB medium for the detection of methicillin-resistant Staphylococcus aureus (MRSA). In total, 102 of 466 specimens yielded MRSA on at least one medium. After incubation for 16-18 h, the sensitivity was 51%, 59%, 47% and 65% on MRSA-ID, CHROMagar MRSA, ORSAB and MRSA Select, respectively, compared with 82%, 75%, 67% and 80%, respectively, after 42 h, and 93%, 95%, 79% and not tested, respectively, following broth enrichment. There were significantly more MRSA colonies on MRSA-Select after 16-18 h than on ORSAB or MRSA ID (p 0.001 and 0.0022, respectively), whereas there were more MRSA colonies after 42 h on MRSA-ID and MRSA-Select than on ORSAB (p 0.0004 and 0.012, respectively). The specificity of the media for identifying MRSA based on the colour of colonies after incubation for 16-18 h was 100%, 99%, 99% and 100%, respectively, compared with 98%, 97%, 98% and 98%, respectively, after 42 h, and 100%, 99%, 100% and not tested, respectively, following broth enrichment. The speed of detection (mean time to report a positive result) was 1.65, 1.72, 2.31 and 1.35 days, respectively. For each of the three media tested following enrichment, the use of an enrichment broth increased the detection rate of MRSA by 16-24%.

[Infective endocarditis and outpatient practice]. / L'endocardite en pratique ambulatoire.

The incidence of infectious endocarditis is fairly stable over the past decades. It is estimated at roughly 3-4 case per patient-year. However, as a consequence of medical progress, Staphylococcus aureus endocarditis has become more prevalent. This is particularly true for health-care associated endocarditis, especially in iv-drug abusers or hemodialysis patients. Mortality (15-20% of patients in the last series) remains high. About 50% of patients undergo surgical treatment, whereas outpatient therapy is more and more frequent for highly selected subgroups of patients without complications and infected with low-risk organims. The present paper reviews in detail the epidemiology of infective endocarditis and discuss in detail the different out-patients therapies.

[Respiratory syncytial virus infections: measures in immunocompromised patients]. / Respiratorisches Syncytial-virus (RSV)-Infektion: Massnahmen beim immunsupprimierten Patienten.

RSV infections are an annually recurring problem of significant morbiditiy and mortality in the immunocompromised host, especially after lung and stem cell transplantation. The effectiveness of hygienic (isolation) and therapeutic (ribavirin, immunoglobulin, pavilizumab) measures depends of high index of suspicion and rapid diagnostic confirmation by the antigen test, PCR and/or culture. Allocation of resources require stratification of measures according to the known or estimated risk for complications.

Evidence for clonal evolution among highly polymorphic genes in methicillin-resistant Staphylococcus aureus.

The evolution of Staphylococcus aureus has been described as predominantly clonal, based on evidence from seven housekeeping genes. We aimed to test if this was also true for more polymorphic genes. In a collection of 60 isolates including major European epidemic methicillin-resistant S. aureus (MRSA) and sporadic MRSA strains, we compared the partial gene sequences of seven housekeeping genes (arcC, aroE, glpF, gmk, pta, tpi, and yqiL), six core adhesion genes (present in all strains) (clfA, clfB, fnbA, map, sdrC, and spa), and four accessory adhesion genes (not present in all strains) (ebpS, fnbB, sdrD, and sdrE). Nucleotide diversity of adhesion genes was 2- to 10-fold higher than genes used for multilocus sequence typing. All genes showed evidence for purifying selection with a weakly reduced level among accessory adhesion genes. Among these highly variable genes, there was no evidence for a difference in molecular evolution between epidemic and sporadic strains. Gene trees constructed from concatenated sequences of housekeeping, core adhesion, and accessory adhesion genes were highly congruent, indicating clonality, despite some evidence for homologous exchange. Further evidence for clonality was found with an overall positive correlation of allelic and nucleotidic divergence for both seven housekeeping genes and six core adhesion genes. However, for small allelic differences that fit the demarcations of clonal complexes (CCs) there was no such correlation, suggesting that recombination occurred. Therefore, despite an overall clonal population structure, recombination between related isolates within CCs might have contributed to S. aureus evolution.

Water disinfection with ozone, copper and silver ions, and temperature increase to control Legionella: seven years of experience in a university teaching hospital.

The efficacy of ozonation, copper-silver ionization and increased temperature in controlling Legionella spp. in the hot water distribution networks of a university hospital was evaluated. Two separate water distribution networks were studied; network 1 which supplies the surgical intensive care units, and network 2 which supplies the medical intensive care units and the emergency room. Network 1 has been disinfected by ozonation since 1995, and network 2 has been disinfected by ionisation since 1999. The hot water temperature was increased from 50 to 65 degrees C in 1998 and 2000 in networks 1 and 2, respectively. Water samples and swabs of the water outlets were cultured for Legionella spp. between four and six times each year, providing data before and after implementation of the disinfection procedures. There was no significant difference in the proportion of samples positive for Legionella spp. after ozonation in network 1 or after ionization in network 2. In both networks, there was a significant reduction in legionella isolates after increasing the hot water temperature to 65 degrees C. Maintaining the hot water temperature above 50 degrees C throughout both networks proved to be the most effective control measure in our hospital.

Influence of bacille Calmette-Guérin vaccination on size of tuberculin skin test reaction: to what size?

BACKGROUND: Previous bacillus Calmette-Guerin (BCG) vaccination can confound the results of a tuberculin skin test (TST). We sought to determine a cutoff diameter of TST induration beyond which the influence of BCG vaccination was negligible in evaluating potential Mycobacterium tuberculosis infection in a population of health care workers with a high vaccination rate and low incidence of tuberculosis. METHODS: From 1991 through 1998, all new employees at the University Hospital of Lausanne, Switzerland, underwent a 2-step TST at entry visit. We also gathered information on demographic characteristics, along with factors commonly associated with tuberculin positivity, including previous BCG vaccination, history of latent M. tuberculosis infection, and predictors for M. tuberculosis infection. RESULTS: Among the 5117 investigated subjects, we found that influence of BCG vaccination on TST results varied across categories of age (likelihood ratio test, 0.0001). Prior BCG vaccination had a strong influence on skin test results of

Risk of Borrelia burgdorferi infection in western Switzerland following a tick bite.

The aim of this study was to define the risk of developing Lyme borreliosis after a tick bite. A survey was conducted from 1993 to 1995 in the western part of Switzerland in a group of patients who presented for treatment of a recent tick bite. Only patients with negative serological tests (enzyme-linked fluorescent assay screening test, and IgG and IgM immunoblots) at the first consultation and for whom a second blood sample was available 2 months later were included in the study. Of the 376 patients included, 266 had no clinical manifestation (group 1) and 110 had a small local cutaneous reaction (<2 cm) (group 2). The tick was available for 160 patients. Seroconversion was observed in 4.5% of 376 patients, 3.4% in group 1 and 7.2% in group 2. Typical erythema migrans, confirmed by seroconversion, was observed in three of 376 (0.8%) patients, while five of 376 (1.3%) patients developed a skin lesion without seroconversion. No other clinical manifestation of Lyme borreliosis was observed among these 376 patients. Borrelia detection in ticks did not correlate significantly with the risk of Lyme borreliosis. In conclusion, the risk of developing Lyme borreliosis in western Switzerland after a tick bite is low, and therefore, prophylactic antibiotics are not required.

Faucets as a reservoir of endemic Pseudomonas aeruginosa colonization/infections in intensive care units.

OBJECTIVE: To evaluate the role of faucets as a reservoir for Pseudomonas aeruginosa colonization/infection of patients hospitalized in intensive care units (ICUs). DESIGN: Prospective epidemiological investigation performed during a nonepidemic period of 1 year. The inner part of the ICU faucets were swabbed for P. aeruginosa. Data were recorded on all patients with at least one culture of a clinical specimens positive for P. aeruginosa. Pulsed-field gel electrophoresis was used to characterize the strains. SETTING: Five ICUs of a university hospital which are supplied by two separate water distribution networks. PATIENTS: During a 1-year period 132 cases were investigated. RESULTS: In 42% of cases (56/132) there were isolates identical to those found in the faucets, with a total of nine different genotypes. Among the nine genotypes isolated from both patients and faucets one of them, the most prevalent, was isolated in the two networks and in 30 cases. The other eight genotypes were recovered almost exclusively from either one (three genotypes in 12 cases) or the other (five genotypes in 12 cases) network and from the patients in the corresponding ICUs. CONCLUSIONS: These results suggest that the water system of the ICUs was the primary reservoir of patient's colonization/infection with P. aeruginosa in a substantial proportion of patients, although the exact mode of acquisition could not be determined.

Molecular epidemiology of predominant clones and sporadic strains of methicillin resistant Staphylococcus aureus in Switzerland and comparison with European epidemic clones.

OBJECTIVE: To assess the molecular epidemiology and risk factors of predominant clones and sporadic strains of methicillin-resistant Staphylococcus aureus (MRSA) in Swiss hospitals and to compare them with European strains of epidemic clones. MATERIAL AND METHODS: One-year national survey of MRSA cases. Analysis of epidemiological and molecular typing data (PFGE) of MRSA strains. RESULTS: In 1997, 385 cases of MRSA were recorded in the five Swiss university hospitals and in 47 community hospitals. Half of the cases were found in Geneva hospitals where MRSA was already known to be endemic. Molecular typing of 288 isolates (one per case) showed that 186 (65%) belong to four predominant clones, three of which were mostly present in Geneva hospitals. In contrast, the fourth clone (85 cases) was found in 23 hospitals (in one to 16 cases per hospital). The remaining 35% of the strains were clustered into 62 pulsed field gel electrophoresis types. They accounted for one to five patients per hospital and were defined as sporadic. Multivariate analysis revealed no independent risk factors for harboring a predominant versus a sporadic strain, except that transfer from a foreign hospital increases the risk of harboring a sporadic strain (OR, 42; 95% CI, 5-360). CONCLUSION: While cases with predominant clones were due to the local spread of these clones, most sporadic cases appear to be due to the continuous introduction of new strains into the country. With the exception of a transfer from a hospital outside Switzerland, no difference in the clinical or epidemiological characteristics was observed between patients harboring a predominant clone and those with a sporadic strain.