Patient-level analysis of incident vancomycin-resistant enterococci colonization and antibiotic days of therapy.

Vancomycin-resistant enterococci (VRE) infections are a public health threat associated with increased patient mortality and healthcare costs. Antibiotic usage, particularly cephalosporins, has been associated with VRE colonization and VRE bloodstream infections (VRE BSI). We examined the relationship between antimicrobial usage and incident VRE colonization at the individual patient level. Prospective, weekly surveillance was undertaken for incident VRE colonization defined by negative admission but positive surveillance swab in a medical intensive care unit over a 17-month period. Antimicrobial exposure was quantified as days of therapy (DOT)/1000 patient-days. Multiple logistic regression was used to analyse incident VRE colonization and antibiotic DOT, controlling for demographic and clinical covariates. Ninety-six percent (1398/1454) of admissions were swabbed within 24 h of intensive care unit (ICU) arrival and of the 380 patients in the ICU long enough for weekly surveillance, 83 (22%) developed incident VRE colonization. Incident colonization was associated in bivariate analysis with male gender, more previous hospital admissions, longer previous hospital stay, and use of cefepime/ceftazidime, fluconazole, azithromycin, and metronidazole (P < 0·05). After controlling for demographic and clinical covariates, metronidazole was the only antibiotic independently associated with incident VRE colonization (odds ratio 2·0, 95% confidence interval 1·2-3·3, P < 0·009). Our findings suggest that risk of incident VRE colonization differs between individual antibiotic agents and support the possibility that antimicrobial stewardship may impact VRE colonization and infection.

Pneumocystis pneumonia in hospitalized patients: a detailed examination of symptoms, management, and outcomes in human immunodeficiency virus (HIV)-infected and HIV-uninfected persons.

BACKGROUND: Pneumocystis jirovecii pneumonia (PCP) is a life-threatening infection for immunocompromised individuals. Robust data and clear guidelines are available for prophylaxis and treatment of human immunodeficiency virus (HIV)-related PCP (HIV-PCP), yet few data and no guidelines are available for non-HIV-related PCP (NH-PCP). We postulated that prevention and inpatient management of HIV-PCP differed from NH-PCP. METHODS: We performed a retrospective case review of all pathologically confirmed cases of PCP seen at the University of Alabama Medical Center from 1996 to 2008. Data on clinical presentation, hospital course, and outcome were collected using a standardized data collection instrument. Bivariate analysis compared prophylaxis, adjunctive corticosteroids, and clinical outcomes between patients with HIV-PCP and NH-PCP. RESULTS: Our analysis of the cohort included 97 cases of PCP; 65 HIV and 32 non-HIV cases. Non-HIV cases rarely received primary prophylaxis (4% vs. 38%, P = 0.01) and received appropriate antibiotics later in the course of hospitalization (5.2 days vs. 1.1 days, P < 0.005). Among transplant patients, NH-PCP was diagnosed a mean of 1066 days after transplantation and most patients were on low-dose corticosteroids (87%) at the time of disease onset. No significant differences in adjunctive corticosteroid use (69% vs. 77%, P = 0.39) and 90-day mortality (41% vs. 28%, P = 0.20) were detected. CONCLUSIONS: Patients who have undergone organ or stem cell transplant remain at risk for PCP for many years after transplantation. In our cohort, patients who developed NH-PCP were rarely given prophylaxis, and initiation of appropriate antibiotics was significantly delayed compared to cases of HIV-PCP. Medical providers should be aware of the ongoing risk for NH-PCP, even late after transplantation, and consider more aggressive approaches to both prophylaxis and earlier empirical therapy for PCP.

Observational study of the epidemiology and outcomes of vancomycin-resistant Enterococcus bacteraemia treated with newer antimicrobial agents.

Vancomycin-resistant Enterococcus bloodstream infections (VRE-BSI) are a growing problem with few clinical trials to guide therapy. We conducted a retrospective study of management and predictors of mortality for VRE-BSI at a tertiary-care centre from January 2005 to August 2008. Univariate and multivariable analyses examined the relationship of patient characteristics and antibiotic therapy with 30-day all-cause mortality. Rates of VRE-BSI increased from 0·06 to 0·17 infections/1000 patient-days (P=0·03). For 235 patients, 30-day mortality was 34·9%. Patients were primarily treated with linezolid (44·2%) or daptomycin (36·5%). Factors associated with mortality were haemodialysis [odds ratio (OR) 3·2, 95% confidence interval (CI) 1·6-6·3, P=0·007], mechanical ventilation (OR 3·7, 95% CI 1·3-10·4, P=0·01), and malnutrition (OR 2·0, 95% CI 1·0-4·0, P=0·046). Use of linezolid, but not daptomycin (P=0·052) showed a trend towards an association with survival. In conclusion, VRE-BSI is a growing problem, associated with significant 30-day mortality. Multiple factors were associated with poor outcomes at our hospital.

Improving molecular detection of fungal DNA in formalin-fixed paraffin-embedded tissues: comparison of five tissue DNA extraction methods using panfungal PCR.

DNA extraction from formalin-fixed paraffin-embedded (FFPE) tissues is difficult and requires special protocols in order to extract small amounts of DNA suitable for amplification. Most described methods report an amplification success rate between 60 and 80%; therefore, there is a need to improve molecular detection and identification of fungi in FFPE tissue. Eighty-one archived FFPE tissues with a positive Gomori methenamine silver (GMS) stain were evaluated using five different commercial DNA extraction kits with some modifications. Three different panfungal PCR assays were used to detect fungal DNA, and two housekeeping genes were used to assess the presence of amplifiable DNA and to detect PCR inhibitors. The sensitivities of the five extraction protocols were compared, and the quality of DNA detection (calculated for each kit as the number of housekeeping gene PCR-positive samples divided by the total number of samples) was 60 to 91% among the five protocols. The efficiencies of the three different panfungals used (calculated as the number of panfungal-PCR-positive samples divided by the number of housekeeping gene PCR-positive samples) were 58 to 93%. The panfungal PCR using internal transcribed spacer 3 (ITS3) and ITS4 primers yielded a product in most FFPE tissues. Two of the five DNA extraction kits (from TaKaRa and Qiagen) showed similar and promising results. However, one method (TaKaRa) could extract fungal DNA from 69 of the 74 FFPE tissues from which a housekeeping gene could be amplified and was also cost-effective, with a nonlaborious protocol. Factors such as sensitivity, cost, and labor will help guide the selection of the most appropriate method for the needs of each laboratory.

Repetitive extragenic palindromic PCR for study of Streptococcus mutans diversity and transmission in human populations.

Pulsed-field gel electrophoresis (PFGE) is considered the "gold standard" for molecular epidemiological study. Repetitive extragenic palindromic PCR (rep-PCR) is less time-consuming and more suitable for analyzing large numbers of bacterial strains in human populations. PFGE and rep-PCR provide comparable genotyping results for investigating Streptococcus mutans diversity and transmission.

Emergence of USA300 MRSA in a tertiary medical centre: implications for epidemiological studies.

Community-associated meticillin-resistant Staphylococcus aureus (CA-MRSA) has become a major pathogen, particularly in outbreaks of skin and soft-tissue infection (SSTI). A preliminary study conducted at our institution in 2004 revealed that up to 45% of inpatient and 70% of outpatient MRSA isolates tested were the USA300 genotype. In this report, we used pulsed-field gel electrophoresis (PFGE) in a retrospective analysis to determine the time when CA-MRSA USA300 moved from the community to the inpatient population. During the five-year period 2000 to 2004, unique MRSA isolates (N=253) were selected from inpatients in surgical and medical intensive care units, the general hospital population and outpatients. The most common PFGE types found in all populations from 2000 to 2003 were USA100, USA200 and USA600. USA300 was absent from all inpatients from 2000 to 2003 and only sporadic numbers found in the outpatient group. However, in 2004 the USA300 strain emerged in both outpatient and hospitalised patients. There was no difference in the distribution of USA300 between ICUs and the general inpatient population. The emergence of CA-MRSA has resulted in a shift of the MRSA strains that are implicated in healthcare-associated infections in our institution. This has been a recent development that has implications as to the use of PFGE to determine transmission of MRSA in the inpatient setting. Further evaluation of these data in the context of the epidemiology of these infections is needed to determine if more discriminatory approaches to typing will be required for monitoring the spread of the more virulent CA-MRSA phenotype within the inpatient population.

IL-1-induced murine osteoblast IL-6 production is mediated by the type 1 IL-1 receptor and is increased by 1,25 dihydroxyvitamin D3.

IL-1-induced osteoblast IL-6 production represents one possible mechanism by which IL-1 augments bone resorption. In this report, we show that the murine osteoblastic cell line (MC3T3-E1) expresses type 1 IL-1 receptors based on 125I-HrIL1 alpha binding, blocked by type 1 IL-1R antibodies (35F5), and analysis of MC3T3 RNA by reverse transcription (RT)-DNA amplification and Northern analysis. MC3T3 cells do not express detectable type 2 IL-1R mRNA by RT-DNA amplification. IL-1 induces (IL-1 ED50, 0.1 pM) IL-6 production through the type 1 IL-1R as 35F5 antibodies block IL-1-stimulated IL-6 production. Vitamin D3 increases IL-1R expression dose- and metabolite-dependently, with 1,25-(OH)2D3 having the greatest potency, and also enhances IL-1's capacity to stimulate IL-6 production at low IL-1 levels. Both IL-1 and 1,25-(OH)2D3 induce type 1 IL-1R and not type 2 IL-1R upregulation based on ligand binding and RT-DNA amplification. Increased IL-1R expression requires a 5-7-h treatment and is protein/RNA synthesis dependent. These observations imply that IL-1-induced IL-6 production in osteoblasts is mediated by type 1 IL-1Rs and that increased IL-1R expression could play a role in mediating IL-1-induced skeletal responses.

Genotypic characterization of Ureaplasma species by pulsed field gel electrophoresis.

We describe the first use of pulsed field gel electrophoresis to genotype human Ureaplasma species. This technique can distinguish between U. urealyticum and U. parvum, differentiate most of the 14 serovars from one another, and identify differences among clinical isolates of the same serovar.

Catheter ablation of accessory pathways, atrioventricular nodal reentrant tachycardia, and the atrioventricular junction: final results of a prospective, multicenter clinical trial. The Atakr Multicenter Investigators Group.

BACKGROUND: The purpose of this study was to evaluate the safety and efficacy of a temperature-controlled radiofrequency catheter ablation system. METHODS AND RESULTS: The patient population included 1050 patients who had undergone ablation of atrioventricular nodal reentrant tachycardia (AVNRT), an accessory pathway (AP), or the atrioventricular junction (AVJ). Ablation was successful in 996 patients. The probability of success was highest among patients who had undergone ablation of the AVJ, lowest in patients who had undergone ablation of an AP, and in between for patients who had undergone ablation of AVNRT. A major complication occurred in 32 patients. Four variables predicted ablation success (AVJ, AVNRT, or left free wall AP ablation and an experienced center). Four factors predicted arrhythmia recurrence (right free wall, posteroseptal, septal, and multiple APs). Two variables predicted development of a complication (structural heart disease and the presence of multiple targets), and 3 variables predicted an increased risk of death (heart disease, lower ejection fraction, and AVJ ablation). CONCLUSIONS: These findings may serve as a guide to clinicians considering therapeutic options in patients who are candidates for ablation.

Usefulness of follow-up electrophysiologic study and event monitoring after successful radiofrequency catheter ablation of supraventricular tachycardia. Atakr Multicenter Investigators Group.

We assessed the usefulness of routine follow-up electrophysiologic studies after successful catheter ablation for supraventricular tachycardia and the role of event monitoring as an alternative modality in 310 patients at 11 centers using an investigational catheter ablation system with closed-loop temperature control. A routine follow-up electrophysiologic study between 1 and 3 months after ablation was required as part of the study protocol, and patients developing palpitations after ablation were encouraged to use event monitors. Recurrence of the initially targeted arrhythmia developed in 23 patients (7.4%) at a mean of 1.5 +/- 1.5 months after ablation. However, only 2 of these 23 recurrences were discovered by routine follow-up electrophysiologic study in asymptomatic patients (both with concealed accessory pathways); in the remaining 21 patients a positive follow-up electrophysiologic study was heralded by either recurrent symptoms, documented recurrent supraventricular tachycardia, and/or preexcitation on the electrocardiogram. Eighteen patients complained of palpitations after ablation and received an event monitor, which correctly diagnosed another cause of palpitations and ruled out recurrence of the ablated arrhythmia in 8 patients. Thus, the combination of clinical follow-up and event monitoring appears to be an effective alternative to routine follow-up electrophysiologic studies after catheter ablation of supraventricular tachycardia.

Reduction of vancomycin-resistant enterococcal infections by limitation of broad-spectrum cephalosporin use in a trauma and burn intensive care unit.

Both vancomycin and third-generation cephalosporin use are believed to contribute to a rise in vancomycin-resistant enterococci (VRE) infections. In 1998, the largest number of VRE infections in our hospital occurred in the trauma/burn intensive care unit (TBICU), accounting for nearly 20% of hospital infections. In an attempt to control the VRE infection rate, antibiotic protocols for prophylaxis, empiric, and definitive therapy were initiated during the final quarter of 1998 to minimize cephalosporin use by the introduction of piperacillin/tazobactam. Therefore, we undertook a study of the VRE infection rate for the TBICU in relation to vancomycin, piperacillin/tazobactam, piperacillin, third-generation cephalosporin, and total cephalosporin use before and after efforts to limit cephalosporins. These data were compared to those in the medical and surgical intensive care units. During 1998, seven VRE infections occurred in the TBICU. Following initiation of antibiotic protocols, one case of VRE infection occurred in the subsequent month and no cases in the 17 months since. The decrease in the VRE infection rate corresponded with a significant increase in the use of piperacillin/tazobactam and a reduction in third-generation and total cephalosporin use. In contrast, cephalosporin use in the medical and surgical intensive care units remains significantly higher than in the TBICU, and neither unit has had a reduction in their VRE infection rates.

Comparison of two rapid latex agglutination tests for detection of cryptococcal capsular polysaccharide.

The Murex Cryptococcus Test was compared with the Cryptococcal Antigen Latex Agglutination System (CALAS) for detecting cryptococcal polysaccharide in 173 cerebrospinal fluid (CSF) specimens and 117 serum samples with 99% and 97% concordance, respectively. Eighteen CSF samples and 17 serum samples were positive in both assays, and 249 were negative. The sensitivity and specificity of the Murex relative to the CALAS were 90% and 100%, respectively, for CSF, and 81% and 100%, respectively, for serum. Six discrepancies were arbitrated by retesting, using a third analytic method, review of other laboratory and clinical data, or both. The reaction in 1 CSF specimen was considered false positive by the CALAS, and the reactions in 2 serum samples were false negatives by the Murex. For 3 patients with previous cryptococcal meningitis but no active disease, only the CALAS detected antigen, suggesting that the Murex has less analytic sensitivity in this context. Titer differences dictate that direct comparisons between the 2 tests are not feasible. There were no false-positive reactions in limited testing with either method using specimens from patients with concurrent noncryptococcal infections or in rheumatoid factor-positive serum samples. Infections caused by Cryptococcus neoformans serotypes A or AD were detected equally by both assays. Based on our study, we have elected to continue to use the CALAS for routine testing for cryptococcal antigen.

Association rules and data mining in hospital infection control and public health surveillance.

OBJECTIVES: The authors consider the problem of identifying new, unexpected, and interesting patterns in hospital infection control and public health surveillance data and present a new data analysis process and system based on association rules to address this problem. DESIGN: The authors first illustrate the need for automated pattern discovery and data mining in hospital infection control and public health surveillance. Next, they define association rules, explain how those rules can be used in surveillance, and present a novel process and system--the Data Mining Surveillance System (DMSS)--that utilize association rules to identify new and interesting patterns in surveillance data. RESULTS: Experimental results were obtained using DMSS to analyze Pseudomonas aeruginosa infection control data collected over one year (1996) at University of Alabama at Birmingham Hospital. Experiments using one-, three-, and six-month time partitions yielded 34, 57, and 28 statistically significant events, respectively. Although not all statistically significant events are clinically significant, a subset of events generated in each analysis indicated potentially significant shifts in the occurrence of infection or antimicrobial resistance patterns of P. aeruginosa. CONCLUSION: The new process and system are efficient and effective in identifying new, unexpected, and interesting patterns in surveillance data. The clinical relevance and utility of this process await the results of prospective studies currently in progress.

Association between antecedent intravenous antimicrobial exposure and isolation of vancomycin-resistant enterococci.

Vancomycin-resistant enterococci (VRE) have become important causes of nosocomial infections. This study evaluated the association between a variety of intravenous antimicrobial exposures and the isolation of VRE using two control groups: (1) a vancomycin-susceptible enterococci (VSE) group, to assess factors associated with development of VRE, and (2) a nonenterococci control group, to assess factors associated with positive cultures for enterococci without regard to vancomycin resistance. After adjusting for the effect of other antimicrobials, time at risk, and patient morbidity, compared to vancomycin-susceptible enterococci controls, exposures to imipenem (OR = 4.9, 95% CI = 1.6-14.1) and ceftazidime (OR = 2.6, 95% CI = 1.1-6.1) were significant predictors of VRE. When compared to nonenterococci controls, exposures to ampicillin (OR = 20.1, 95% CI = 1.5-263.1) and imipenem (OR = 5.1, 95% CI = 1.5-17.1) were significantly associated with VRE. Neither piperacillin nor vancomycin was associated with VRE compared to either control group. This study offers further evidence that the replacement of broad-spectrum cephalosporins by extended-spectrum penicillins, specifically piperacillin, may be effective in reducing VRE.

Direct bacterial identification from positive BacT/Alert blood cultures using MicroScan overnight and rapid panels.

Studies were conducted on a method of direct inoculation of MicroScan overnight and rapid panels from positive BacT/Alert blood culture bottles containing standard aerobic media to determine the correlation with inoculation of the corresponding panels with a standardized bacterial suspension obtained following subculture to agar. For Gram-negative organisms, 122 of 127 (96%) overnight panels and 85 of 118 (72%) rapid panels showed complete agreement with the standard method for species identification. Highest concordance (99%) occurred with Enterobacteriaceae inoculated directly into overnight panels. For Gram-positive organisms, 70 of 85 (82%) overnight panels and 45 of 86 (52%) rapid panels showed complete agreement. These findings suggest that direct inoculation of Gram-negative overnight MicroScan panels yields results most comparable to standard methods when Enterobacteriaceae are detected and allows reporting of results 18 to 24 h sooner. Direct inoculation of Gram-positive overnight or rapid panels and Gram-negative rapid panels from this blood culture medium did not yield acceptable identification results and is not recommended.

Trends in frequency and susceptibilities of Candida glabrata bloodstream isolates at a university hospital.

The frequency of isolation and antifungal susceptibility patterns to fluconazole and itraconazole were determined for 166 Candida glabrata isolates causing bloodstream infection at a single institution from 1995-2000. Findings demonstrated a trend of increasing resistance to itraconazole among the isolates, but no trend in resistance to fluconazole. The frequency of C. glabrata isolates among all blood culture isolates of Candida spp. causing bloodstream infection remained stable during the study period and ranged from 18-31%.

Detection of oxacillin-resistance in Staphylococcus aureus by MicroScan MIC panels in comparison to four other methods.

Two hundred fifty-two isolates of Staphylococcus aureus were tested for oxacillin susceptibility by MicroScan Gram positive overnight and rapid MIC panels. Results were compared with nonautomated methods including disk diffusion, MRSA Crystal ID, and Etests using MRSA Screen Agar as reference. One hundred sixty-nine isolates (67.1%) were oxacillin-susceptible and 83 (32.9%) were resistant. All methods agreed for 234 (92.9%) isolates. Very major error rates were 1.2% for disk diffusion, 3.6% for Etest, and 0 for all other methods. Major error rates were 5.3% for MicroScan overnight panels, 3% for rapid panels, 2.4% for disk diffusion, 1.2% for Etest, and 0.6% for MRSA Crystal ID. Nine oxacillin-susceptible isolates with borderline MICs and discrepant results for 1 or more methods were tested for the mec A gene and all were negative. Each was susceptible to beta lactam/beta lactamase inhibitor combinations, suggesting that false resistance may have been due to excessive beta lactamase production. Oxacillin-resistant S. aureus with borderline MICs determined by MicroScan should be confirmed by an alternate method. The most practical and cost-effective means among those we tested is the MRSA Screen Agar.

Genotypic investigation of multidrug-resistant Acinetobacter baumannii infections in a medical intensive care unit.

Multi-resistant Acinetobacter baumannii isolates obtained from 13 hospitalized patients over a six-month period were evaluated. One patient had an isolate susceptible only to imipenem; the next three had isolates susceptible to imipenem and ampicillin/sulbactam; the next six patients had isolates which were susceptible to imipenem, amikacin, and ampicillin/sulbactam; while the final three patients had isolates which were susceptible to imipenem and ampicillin/sulbactam. Ten patients died, five within 10 days of a positive culture. Five of six patients with bacteraemia succumbed to the infection. DNA extracted from all isolates was amplified by polymerase chain reaction using four random primers (RAPD). The resulting band patterns were compared and strains identified. In addition, all isolates were biotyped. RAPD analysis and biotyping showed that there were two distinct strains involved. The first four patients were infected with one strain (genotype ¿A', biotype 9), the subsequent nine patients were infected with a second strain (genotype ¿B', biotype 1). These results suggested that there was patient-to-patient spread of strains. Institution of, and strict adherence to, isolation procedure and other infection control practices controlled the spread of infection. These data emphasize the need for active surveillance for multidrug-resistant organisms in critically ill patients, and the value of molecular typing of strains in a hospital setting to investigate spread of infection.

Vancomycin-resistant enterococci: 15 years and counting.

We review the history of vancomycin-resistant enterococci (VRE) and propose a causal model illustrating the roles of exposure to VRE reservoirs, patient characteristics, antimicrobial exposure, and prevalence of VRE in the progression from potential VRE reservoirs to active disease in hospitalized patients. Differences in VRE colonization and VRE infection are discussed with respect to hospital surveillance methodology and implications for interventions. We further document clonal transmission of VRE in a large, urban, teaching hospital and demonstrate VRE susceptibility to a wide array of antimicrobial agents. This model can guide the identification of mutable factors that are focal points for intervention.

Pathology information systems: data mining leads to knowledge discovery.

Information systems in pathology provide opportunities for pathologists and clinical laboratory scientists to impact both clinical care and modern research agendas. The paradigm shift in health care from individualized care to population-based and standardized delivery systems has created both of these opportunities. In research, pathology information systems can provide key databases for health services research and new informatics-based approaches to database research. The latter is characterized by utilization of pathology databases for data mining to discover new patterns that provide new knowledge. The multidisciplinary knowledge discovery and data mining program at the University of Alabama at Birmingham focuses on this health care application, which has the potential to make a major impact on health care research and delivery.