Marta Rodríguez-Créixems

Reassessment of Clostridium difficile Susceptibility to Metronidazole and Vancomycin

ABSTRACT Clostridium difficile is the most frequently identified enteric pathogen in patients with nosocomially acquired, antibiotic-associated diarrhea. The drugs most commonly used to treat diseases associated with C. difficile are metronidazole and vancomycin. Most clinical laboratories assume that all C. difficile isolates are susceptible to metronidazole and vancomycin. We report on the antimicrobial susceptibilities of 415 C. difficile isolates to metronidazole and vancomycin over an 8-year period (1993 to 2000). The overall rate of resistance to metronidazole at the critical breakpoint (16 μg/ml) was 6.3%. Although full resistance to vancomycin was not observed, the overall rate of intermediate resistance was 3.1%. One isolate had a combination of resistance to metronidazole and intermediate resistance to vancomycin. Rates of resistance to metronidazole and vancomycin were higher among isolates from human immunodeficiency virus-infected patients. Molecular typing methods proved the absence of clonality among the isolates with decreased susceptibilities to the antimicrobials tested.

Successful Outcome of Scedosporium apiospermum Disseminated Infection Treated with Voriconazole in a Patient Receiving Corticosteroid Therapy

A disseminated Scedosporium apiospermum infection was diagnosed in a woman with severe asthma and treated with corticosteroids. This fungi is resistant to fluconazole and amphotericin B. The infection was refractory to itraconazole, but responded successfully to voriconazole. A review of the literature is provided.

Molecular diagnosis of infective endocarditis by real-time broad-range polymerase chain reaction (PCR) and sequencing directly from heart valve tissue

Traditionally, infective endocarditis (IE) has been microbiologically diagnosed by blood cultures or serology. However, conventional microbiologic methods do not always provide an etiologic diagnosis. We conducted the current study to evaluate the usefulness of a universal real-time polymerase chain reaction (PCR) of the 16S rRNA gene followed by sequencing for the diagnosis of IE in explanted heart valve tissue (HV) as part of the routine of a clinical microbiology laboratory, and to compare it with conventional culture of blood or HV. We prospectively analyzed 177 HV samples by universal PCR and sequencing: 48 were from 35 patients with definite IE and 129 were from 120 patients without IE. Specific PCR tests were used when necessary to confirm broad-range PCR results. For the 35 patients with IE, all of the HV samples except for 2 from the same patient gave positive PCR results. The microorganisms identified matched those isolated by blood culture in 31 cases. The other 3 patients had negative blood culture IE, but PCR made possible the detection of Tropheryma whipplei, Bartonella quintana, and Streptococcus gallolyticus. For the negative control group, universal PCR was completely negative in 123 of the 129 samples. Sensitivity, specificity, and negative and positive predictive values of this real-time PCR method were 96%, 95.3%, 98.4%, and 88.5%, respectively, for the diagnosis of IE, using the Duke criteria to define IE and using blood culture results to identify etiologic microorganisms. Conventional HV culture correlated poorly with blood cultures and molecular techniques, and frequently represented tissue contamination resulting from valve handling. Our universal PCR method has proved to be more sensitive, specific, and rapid than conventional culture methods, and should therefore be included as a new major Duke criterion for the diagnosis of IE. According to the results of the current study, this technique should be used to supplement blood and HV culture. Conventional HV cultures are frequently responsible for false-positive and false-negative results, and are not always useful to establish the etiology of IE. Abbreviations: HV = heart valve tissue, IE = infective endocarditis, PCR = polymerase chain reaction.

Heterogeneous Vancomycin-Intermediate Susceptibility Phenotype in Bloodstream Methicillin-Resistant Staphylococcus aureus Isolates from an International Cohort of Patients with Infective Endocarditis: Prevalence, Genotype, and Clinical Significance

BACKGROUND The significance of heterogeneous vancomycin-intermediate Staphylococcus aureus (hVISA) is unknown. Using a multinational collection of isolates from methicillin-resistant S. aureus (MRSA) infective endocarditis (IE), we characterized patients with IE with and without hVISA, and we genotyped the infecting strains. METHODS MRSA bloodstream isolates from 65 patients with definite IE from 8 countries underwent polymerase chain reaction (PCR) for 31 virulence genes, pulsed-field gel electrophoresis, and multilocus sequence typing. hVISA was defined using population analysis profiling. RESULTS Nineteen (29.2%) of 65 MRSA IE isolates exhibited the hVISA phenotype by population analysis profiling. Isolates from Oceania and Europe were more likely to exhibit the hVISA phenotype than isolates from the United States (77.8% and 35.0% vs 13.9%; P < .001). The prevalence of hVISA was higher among isolates with a vancomycin minimum inhibitory concentration of 2 mg/L (P = .026). hVISA-infected patients were more likely to have persistent bacteremia (68.4% vs 37.0%; P = .029) and heart failure (47.4% vs 19.6%; P = .033). Mortality did not differ between hVISA- and non-hVISA-infected patients (42.1% vs 34.8%, P = .586). hVISA and non-hVISA isolates were genotypically similar. CONCLUSIONS In these analyses, the hVISA phenotype occurred in more than one-quarter of MRSA IE isolates, was associated with certain IE complications, and varied in frequency by geographic region.

Bloodstream Infections: A Trial of the Impact of Different Methods of Reporting Positive Blood Culture Results

BACKGROUND The impact of how positive blood culture results are reported on the evolution bloodstream infections (BSIs) has not been assessed. METHODS We randomly assigned patients with BSIs into 3 groups: group A (for which physicians received a conventional report), group B (for which physicians received a conventional report and a written alert on the chart with clinical advice), and group C (for which physicians received the above plus oral clinical advice). The adequacy of therapy before and after receipt of the different types of information was assessed. RESULTS Overall, 297 episodes (109 in group A, 99 in group B, and 89 in group C) were studied. Patients who received inadequate treatment before receiving microbiological information had a longer mean (+/-SD) hospital stay (27.2+/-32.4 vs. 19.4+/-15.8 days; P=.017), a higher mean risk of Clostridium difficile-associated diarrhea (8.3% vs. 1.9%; P=.013), a higher mean overall mortality rate (30.8% vs. 19.4%; P=.025), and a higher mean risk of infection-related mortality (23.3% vs. 13.6%; P=.031). After receipt of microbiological reports, recommendations for changes in therapy were issued for patients in groups B (52.3%) and C (53.1%). For groups A, B, and C, the proportions of days on which adequate treatment was received were 66.3%, 92.1%, and 91.2% (P<.001); the mean numbers of defined daily doses of appropriate antibiotic therapy were 16.4, 22.2, and 20.7 (P=.003); the mean durations of hospital stay were 19.8, 23.6, and 24.1 days (P=.761); and the mortality rates during the late period were 12.9%, 15.6%, and 11% (P=.670), respectively. The mean costs of antimicrobials per episode in groups A, B, and C were 580.63, 537.98, and 434.53 (US707.85 dollars, US699.73 dollars, and US529.73 dollars, respectively). CONCLUSIONS Written- or oral-alert reports with clinical advice should complement traditional microbiological reports for patients with BSIs.

Evolution of susceptibilities of Campylobacter spp. to quinolones and macrolides.

Erythromycin, new macrolides, and quinolones are alternatives for the treatment of Campylobacter infections. Concerns related to the emergence of resistance to both groups of drugs have been raised. We studied the evolution of antimicrobial susceptibilities of 275 clinical isolates of microorganisms of the genus Campylobacter isolated in our institution during a 5-year period (1988 to 1992). The microorganisms studied were C. jejuni (n = 230), C. coli (n = 42), and C. fetus (n = 3). The overall resistance rates (determined by the agar dilution method and the recommendations of the National Committee for Clinical Laboratory Standards) were as follows: erythromycin, 2.3%; clarithromycin, 2.3%; azithromycin, 1.9%; ciprofloxacin, 28.5%; norfloxacin, 31%; ofloxacin, 26.3%; and nalidixic acid, 36.8%. The evolution of resistance (percent resistance in 1988 versus percent resistance in 1992) was as follows: erythromycin, 2.6 versus 3.1; clarithromycin, 2.6 versus 3.1; azithromycin, 2.6 versus 3.1; ciprofloxacin, 0 versus 49.5; norfloxacin, 2.6 versus 55.5; ofloxacin, 0 versus 45.6; nalidixic acid, 2.6 versus 56.8. Our data show stable macrolide activity against Campylobacter spp. and the rapid development of quinolone resistance over the last 5 years.

Is the Volume of Blood Cultured Still a Significant Factor in the Diagnosis of Bloodstream Infections

ABSTRACT “The higher the volume of blood cultured the higher the yield of blood cultures” has been a well-accepted dictum since J. A. Washington II performed his classic work. This rule has not been questioned in the era of highly automated blood culture machines, nor has it been correlated with clinical variables. Our objective in this study was to complete a prospective analysis of the relationship between blood volume, the yield of blood cultures, and the severity of clinical conditions in adult patients with suspected bloodstream infections (BSI). During a 6-month period, random samples of blood cultures were weighed to determine the volume of injected blood (weight/density). Overall, 298 patients with significant BSI and 303 patients with sepsis and negative blood cultures were studied. The mean volume of blood cultured in patients with BSI (30.03 ± 14.96 ml [mean ± standard deviation]) was lower than in patients without BSI (32.98 ± 15.22 ml [P = 0.017]), and more episodes of bacteremia were detected with <20 ml (58.9%) than with >40 ml (40.2%) of blood cultured (P = 0.022). When patients were stratified according to the severity of their underlying condition, patients with BSI had higher APACHE II scores, and higher APACHE II scores were related to lower sample volumes (P < 0.001). A multivariate analysis showed that in the group of patients with APACHE II scores of ≥18, higher volumes yielded higher rates of bacteremia (odds ratio, 1.04 per ml of blood; 95% confidence interval, 1.001 to 1.08). We conclude that the higher yield of blood cultures inoculated with lower volumes of blood reflects the conditions of the population cultured. Washingtons dictum holds true today in the era of automated blood culture machines.

Campylobacter bacteremia: clinical characteristics, incidence, and outcome over 23 years.

Campylobacter is a very rare cause of bloodstream infection, although it has been found relatively frequently in patients infected with human immunodeficiency virus (HIV). The impact of highly active antiretroviral therapy (HAART) and new forms of immunosuppression on the incidence of Campylobacter bacteremia has not been sufficiently assessed. In this study we analyzed the incidence and microbiologic and clinical characteristics of Campylobacter bacteremia over 23 years. We reviewed the clinical records of all patients who had Campylobacter bacteremia from 1985 to 2007. Available strains were reidentified using universal polymerase chain reaction (PCR). During the study period, there were 71 episodes of Campylobacter bacteremia in 63 patients (0.24% of all bloodstream infections), and the incidence remained stable (mean, 0.06/1000 admissions per year and 0.47/100,000 inhabitants per year). Median age was 52 years (interquartile range, 31.25-72.5 yr), and 82% of patients were male. The underlying conditions included liver disease (21/64, 32.8%), HIV infection (15/64, 23.4%), malignancy (7/64, 10.9%), solid organ transplantation (2/64, 3%), hypogammaglobulinemia (10/64, 15.6%), and other (18/64, 31.2%). Twelve patients shared more than 1 underlying condition. Campylobacter bacteremia was community acquired in 81% of the episodes. The origin of the bloodstream infection was abdominal (43.5%), primary (26%), or extraintestinal (31%: respiratory 15%, cellulitis 4.8%, urinary 8%, other 3%). C jejuni was recovered in 66% of cases, C fetus in 19%, and C coli in 12%. Universal PCR was performed on 14 available strains. Molecular and conventional identification matched in 8 isolates. In contrast, molecular methods classified as C fetus (n = 2) and C jejuni (n = 1) 3 strains formerly identified only to genus level as Campylobacter species. In another 3 isolates, molecular identification was not consistent with the phenotypic identification (C fetus identified as C jejuni). Complications appeared in 23.9% of patients. Quinolone resistance was observed in 50% of the isolates. Only 37.8% of patients received appropriate empirical therapy. Mortality was 16.4%, although it was higher in HIV-infected patients than uninfected patients (33% vs. 10%; p = 0.04), in cases of hospital-acquired Campylobacter bacteremia compared with community-acquired cases (38.5% vs. 9.4%; p = 0.02), and in the presence of complications compared with patients without complications (100% vs. 0%; p < 0.001). The incidence of recurrence was 5% (3 patients with humoral immunodeficiency). There was a higher proportion of HIV-infected patients among patients with Campylobacter bacteremia in the pre-HAART era (1985-1996) than in the HAART era (1997-2007)-27.5% (11/40) vs. 14.3% (4/28)-although the difference was not statistically significant. Debilitating diseases such as chronic obstructive pulmonary disease emerged as predisposing conditions in the HAART era (0% before HAART era vs. 14.3% in HAART era; p = 0.032). Campylobacter bacteremia is no longer a significant disease of HIV-positive patients on HAART, but often affects other immunocompromised patients as well. Campylobacter bacteremia has an extraintestinal origin in as many as 31% of cases, and humoral immunodeficiency must be sought in patients with recurrent episodes. Quinolones should not be considered for empirical therapy. Abbreviations: AIDS = acquired immunodeficiency syndrome, ARIMA = autoregressive integrated moving average test, BSI = bloodstream infection, COPD = chronic obstructive pulmonary disease, HAART = highly active antiretroviral therapy, HIV = human immunodeficiency virus, IQR = interquartile range, PCR = polymerase chain reaction, rRNA = ribosomal ribonucleic acid.

A Prospective, Randomized, and Comparative Study of 3 Different Methods for the Diagnosis of Intravascular Catheter Colonization

BACKGROUND Demonstration of catheter tip colonization is usually performed by use of Makis semiquantitative technique, although the superiority of quantitative techniques has been claimed on the basis of their purported ability to detect both endoluminal and exoluminal microorganisms. METHODS We prospectively compared Makis semiquantitative technique and the quantitative methods of sonication and vortexing for the detection of colonization of intravascular catheter tips and catheter-related bloodstream infections. All 3 techniques were performed on the tip of each catheter, and the order in which each technique was performed was randomly assigned. RESULTS Of the 1000 catheter tips that were processed, 329 (32.9%) had positive results for at least 1 of the 3 techniques when a breakpoint of >or=100 colony-forming units (cfu)/catheter segment was used for the quantitative techniques and a breakpoint of >or=15 cfu was used for Makis technique. Eighty-two of the catheter tips for which results were positive were from patients with catheter-related bloodstream infections. For each technique, the likelihood of detection decreased progressively depending on the order in which the technique was performed (i.e., second vs. first and third vs. second). The likelihood of detection of catheter colonization for each technique, when the technique was performed first and when 2 breakpoints (>or=100 cfu/catheter segment [criterion B] and >or=1000 cfu/catheter segment [criterion A]) were used for the quantitative techniques and a breakpoint of >or=15 cfu was used for Makis technique, was as follows: 99.1% and 100% for Makis technique, 95.1% and 92.9% for sonication, and 93.1% and 72.8% for vortexing (for criteria B and A, respectively). No inferiority of Makis technique could be demonstrated when results were compared according to whether catheter placement was short term (i.e., <7 days) or long term (i.e., >or=7 days), either for the detection of colonization or for the detection of catheter-related bloodstream infections. CONCLUSIONS According to data from the present study, the quantitative techniques of sonication and vortexing were not superior to Makis technique under the test conditions used. The greater simplicity of Makis semiquantitative technique makes it the procedure of choice for routine work in the microbiology laboratory.

Bloodstream Infections: Evolution and Trends in the Microbiology Workload, Incidence, and Etiology, 1985-2006

Abstract Information available on bloodstream infection (BSI) is usually restricted to short periods of time, certain clinical backgrounds, or specific pathogens, or is just outdated. We conducted the current prospective study of patients with BSI in a 1750-bed teaching hospital to evaluate workload trends and the incidence and etiology of BSI in a general hospital during the last 22 years, including the acquired immunodeficiency syndrome (AIDS) era. The main outcome measures were laboratory workload, trends in incidence per 1000 admissions and per 100,000 population of different microorganisms, and the impact of the human immunodeficiency virus (HIV) epidemic in the period 1985-2006. From 1985 to 2006 we had 27,419 episodes of significant BSI (22,626 patients). BSI incidence evolved from 16.0 episodes to 31.2/1000 admissions showing an annual increase of 0.83 episodes/1000 admissions (95% confidence interval, 0.61-1.05; p < 0.0001). The evolution of the incidence per 1000 admissions and per 100,000 population of different groups of microorganisms was as follows: Gram positives 8.2 to 15.7/1000 admissions and 66.8 to 138.3/100,000 population; Gram negatives 7.8 to 16.2/1000 admissions and 63.5 to 141.9/100,000 population; anaerobes 0.5 to 1.3/1000 admissions and 4.1 to 11.7/100,000 population; and fungi 0.2 to 1.5/1000 admissions and 1.7 to 12.5/100,000 population. All those differences were statistically significant. We observed the emergence of multiresistant Gram-positive and Gram-negative microorganisms. At least 2484 episodes of BSI (9.1%) occurred in 1822 patients infected with HIV. The incidence of BSI in HIV-infected patients increased from 1985 and reached a peak in 1995 (17.6% of BSI). Since 1995, the decrease was continuous, and in 2006 only 3.9% of all BSI episodes occurred in HIV-positive patients in our institution. We conclude that the BSI workload has increased in modern microbiology laboratories. Gram-positive pathogens have overtaken other etiologic agents of BSI. Our observation shows the remarkable escalation of some resistant pathogens, and the rise and relative fall of BSI in patients with HIV. Abbreviations: AIDS = acquired immunodeficiency syndrome, BSI = bloodstream infection, ESBL = extended-spectrum beta-lactamase, HAART = highly active antiretroviral therapy, HIV = human immunodeficiency virus, MRSA = methicillin-resistant Staphylococcus aureus.