Stephen M. Brecher

Escherichia coli Sequence Type 131 (ST131) Subclone H30 as an Emergent Multidrug-Resistant Pathogen Among US Veterans

BACKGROUND Escherichia coli sequence type 131 (ST131), typically fluoroquinolone-resistant (FQ-R) and/or extended-spectrum β-lactamase (ESBL)-producing, has emerged globally. We assessed its prevalence and characteristics among US veterans. METHODS In 2011, 595 de-identified E. coli clinical isolates were collected systematically within 3 resistance groups (FQ-susceptible [FQ-S], FQ-R, and ESBL-producing) from 24 nationally distributed Veterans Affairs Medical Centers (VAMCs). ST131 and its H30 subclone were detected by polymerase chain reaction and compared with other E. coli for molecular traits, source, and resistance profiles. RESULTS ST131 accounted for 78% (184/236) of FQ-R and 64.2% (79/123) of ESBL-producing isolates, but only 7.2% (17/236) of FQ-S isolates (P < .001). The H30 subclone accounted for ≥95% of FQ-R and ESBL-producing, but only 12.5% of FQ-S, ST131 isolates (P < .001). By back-calculation, 28% of VAMC E. coli isolates nationally represented ST131. Overall, ST131 varied minimally in prevalence by specimen type, inpatient/outpatient source, or locale; was the most prevalent ST, followed distantly by ST95 and ST12 (13% each); and accounted for ≥40% (β-lactams), >50% (trimethoprim-sulfamethoxazole , multidrug), or >70% (ciprofloxacin, gentamicin) of total antimicrobial resistance. FQ-R and ESBL-producing ST131 isolates had higher virulence scores than corresponding non-ST131 isolates. ST131 pulsotypes overlapped extensively among VAMCs. CONCLUSIONS Among US veterans, ST131, primarily its H30 subclone, accounts for most antimicrobial-resistant E. coli and is the dominant E. coli strain overall. Possible contributors include multidrug resistance, extensive virulence gene content, and ongoing transmission. Focused attention to ST131, especially its H30 subclone, could reduce infection-related morbidity, mortality, and costs among veterans.

Direct Identification of Staphylococcus aureus from Positive Blood Culture Bottles

ABSTRACT Fluorescence in situ hybridization (FISH) using peptide nucleic acid (PNA) probes targeting Staphylococcus aureus 16S rRNA is a novel method for direct identification of S. aureus from positive blood culture bottles. The test (S. aureus PNA FISH) is performed on smears made directly from positive blood culture bottles with gram-positive cocci in clusters (GPCC) and provides results within 2.5 h. A blinded comparison of S. aureus PNA FISH with standard identification methods was performed in collaboration with eight clinical microbiology laboratories. A total of 564 routine blood culture bottles positive for GPCC recovered from both aerobic and anaerobic media from three different manufacturers (ESP, BACTEC, and BacT/Alert) were included in the study. The sensitivity and specificity of S. aureus PNA FISH were 100% (57 of 57) and 99.2% (116 of 117), respectively, with 174 GPCC-positive ESP blood culture bottles, 98.5% (67 of 68) and 98.5% (129 of 131), respectively, with 200 GPCC-positive BACTEC blood culture bottles, and 100% (74 of 74) and 99.1% (115 of 116), respectively, with 190 GPCC-positive BacT/Alert blood culture bottles. It is concluded that S. aureus PNA FISH performs well with commonly used continuously monitoring blood culture systems.

Clinical Correlation of the CLSI Susceptibility Breakpoint for Piperacillin- Tazobactam against Extended-Spectrum-β-Lactamase-Producing Escherichia coli and Klebsiella Species

ABSTRACT We assessed infections caused by extended-spectrum-β-lactamase-producing Escherichia coli or Klebsiella spp. treated with piperacillin-tazobactam to determine if the susceptibility breakpoint predicts outcome. Treatment was successful in 10 of 11 nonurinary infections from susceptible strains and in 2 of 6 infections with MICs of >16/4 μg/ml. All six urinary infections responded to treatment regardless of susceptibility.

Laboratory Diagnosis of Clostridium difficile Infections: There Is Light at the End of the Colon

Single molecular or multistep assays (glutamate dehydrogenase, toxin A/B, ± molecular) are recommended for the diagnosis of CDI in patients with clinically significant diarrhea. Rapid and accurate tests can improve resource allocations and improve patient care. Enzyme immunoassay (EIA) for toxins A/B is too insensitive for use as a stand-alone assay. This guideline will examine the use of molecular tests and multitest algorithms for the diagnosis of Clostridium difficile infection (CDI). These new tests, alone or in a multistep algorithm consisting of >1 assay, are more expensive than the older EIA assays; however, rapid and accurate testing can save money overall by initiating appropriate treatment and infection control protocols sooner and by possibly reducing length of hospital stay. We recommend testing only unformed stool in patients with clinically significant diarrhea by a molecular method or by a 2- to 3-step algorithm.

The Clinical Predictive Value (or Lack Thereof) of the Results of In Vitro Antimicrobial Susceptibility Tests

Estimates of antibacterial activity can be determined using a wide variety of different in vitro methods. These include manual methods such as disk diffusion, broth microdilution, and Etests or instrument-based methods such as Vitek (bioMerieux, Durham, NC), Phoenix (BD, Franklin Lakes, NJ),

Variation and persistence of methicillin-resistantStaphylococcus aureus strains among individual patients over extended periods of time

To determine the strain variation and persistence among isolates of methicillin-resistantStaphylococcus aureus (MRSA) cultured from patients with colonization over extended time spans, pulsed-field gel electrophoresis was used to analyze the isolates from 47 patients for whom at least twomecA-positive isolates collected a minimum of six months apart were available. For 22 (47 %) patients, the isolates represented multiple distinct strains ofStaphylococcus aureus, while 20 (43 %) patients had only a single strain detected; five (11 %) patients had similar, genetically related isolates. MRSA were frequently associated with mucocutaneous abnormalities; 29 (62 %) patients had focal cutaneous defects, and ten (21 %) had chronic dermatitis. Multiple strains of MRSA were detected more frequently than single strains among patients in whom the initial focus of MRSA resolved clinically and another mucocutaneous defect subsequently developed compared to patients with clinically persistent foci (11/15 versus 9/23, respectively; p=0.05, Fishers exact test). Among the 21 patients in this series for whom isolates cultured within a two-month time span were available, there were seven (33 %) patients with multiple strains of MRSA, including one patient with polyclonal bacteremia. In summary, patients with long-term MRSA colonization often have several different strains of MRSA, which typically change over time in association with removal or resolution of a colonized focus and the recurrence of mucocutaneous defects.

Extranasal Methicillin-Resistant Staphylococcus aureus Colonization at Admission to an Acute Care Veterans Affairs Hospital

OBJECTIVE To evaluate the prevalence of and risk factors for extranasal methicillin-resistant Staphylococcus aureus (MRSA) colonization and its relationship to nasal colonization among veterans hospitalized for acute care. DESIGN Prospective observational study. SETTING Veterans Affairs (VA) acute care hospital in Boston, Massachusetts. PATIENTS Convenience sample of 150 patients hospitalized within the previous 36 hours and screened for nasal MRSA who were not known to have an active MRSA infection or MRSA isolates recovered from a wound during the past 12 months. METHODS Potential risk factors for MRSA colonization were assessed, and oropharynx, axilla, hand, perirectal, wound, and catheter insertion site samples were obtained for culture. MRSA was identified in chromogenic agar and confirmed by use of routine culture techniques. Nasal MRSA colonization was detected by means of polymerase chain reaction (PCR). RESULTS Nasal swab samples analyzed by use of PCR yielded results positive for MRSA in 16 (11%) of 150 patients. Extranasal cultures yielded positive results for 3 (2%) of 134 patients who tested negative for nasal MRSA colonization and for 9 (56%) of 16 patients who tested positive for nasal MRSA colonization (odds ratio [OR], 56.1 [95% confidence interval {CI}, 12.4-254.6]; p < .001). The oropharynx was the most commonly colonized extranasal site (10 patients [7%]). Independent risk factors for extranasal MRSA colonization included nasal MRSA colonization (OR, 66.9 [95% CI, 11.8-379.7]; P < .001) and end-stage hepatic disease (OR, 98.5 [95% CI, 3.1-3,112.4]; P = .01). CONCLUSIONS Extranasal MRSA colonization is infrequent among veterans admitted for acute care to VA Boston Healthcare System. Extranasal MRSA colonization was strongly associated with nasal MRSA colonization, which suggests that the VA MRSA Prevention Initiative is not missing a large number of MRSA-colonized patients by focusing on nasal-only screening.

Sampling Variability in the Microbiological Evaluation of Expectorated Sputa and Endotracheal Aspirates

ABSTRACT A five-center study was conducted with the aim of determining how reproducibly expectorated sputa and tracheal aspirates could be sampled when preparing Gram-stained smears and inoculating cultures. With both specimen types, excessive variation was noted among Gram stain results obtained from replicate smears. Less variation was noted among culture results, especially with tracheal aspirates.

WSES guidelines for management of Clostridium difficile infection in surgical patients

In the last two decades there have been dramatic changes in the epidemiology of Clostridium difficile infection (CDI), with increases in incidence and severity of disease in many countries worldwide. The incidence of CDI has also increased in surgical patients. Optimization of management of C difficile, has therefore become increasingly urgent. An international multidisciplinary panel of experts prepared evidenced-based World Society of Emergency Surgery (WSES) guidelines for management of CDI in surgical patients.

Clostridium difficile Ribotype Diversity at Six Health Care Institutions in the United States

ABSTRACT Capillary-based PCR ribotyping was used to quantify the presence/absence and relative abundance of 98 Clostridium difficile ribotypes from clinical cases of disease at health care institutions in six states of the United States. Regionally important ribotypes were identified, and institutions in close proximity did not necessarily share more ribotype diversity than institutions that were farther apart.