Kanchana Manickam

UF-1000i flow cytometry is an effective screening method for urine specimens.

This study was undertaken to evaluate the UF-1000i™ (UF) flow cytometer to count urine constituents including bacteria. The objective was to screen urine samples and determine what white blood cell (WBC) and/or bacteria screening criteria would minimize the number of specimens cultured yet ensuring that all true positives were cultured. UF screening and culture on CHROMagar™ Orientation (CO) medium were performed on 2496 specimens. Various combinations of WBC/bacterial counts were assessed as screening criteria and correlated with significant growth on CO medium. A bacterial count of ≥20 from UF gave an overall screening sensitivity of 92.6%, allowing 35% of specimens to be screened out and not cultured. The sensitivity was 99.2% and 85.0% for Gram-negative and Gram-positive organisms, respectively, using the same bacterial count. Our study indicated that UF was a simple, rapid, and reliable method for urine screening when the bacterial count of ≥20 was used as the sole screening criterion.

First report of Actinobaculum schaalii urinary tract infection in North America

A recurrent urinary tract infection with Actinobaculum schaalii, a fastidious, facultative anaerobic, and emerging pathogen, is described. Diagnosis was delayed when routine urine cultures were initially performed yielding recurrently negative results. Resolution of symptoms occurred after anaerobic cultures were done to allow organism isolation, identification, and appropriate antimicrobial treatment.

Reductions in Workload and Reporting Time by Use of Methicillin-Resistant Staphylococcus aureus Screening with MRSASelect Medium Compared to Mannitol-Salt Medium Supplemented with Oxacillin

ABSTRACT Methicillin-resistant Staphylococcus aureus (MRSA) is a significant pathogen in both nosocomial and community settings, and screening for carriers is an important infection control practice in many hospitals. In this retrospective study, we demonstrate that the implementation of an MRSA screening protocol using a selective chromogenic medium (MRSASelect) reduced the workload for this screening test by 63.7% overall and by 12.6% per specimen and reduced the turnaround time for reporting by an average of 1.33 days for all MRSA screening specimens, 1.97 days for MRSA-positive specimens, and 1.3 days for MRSA-negative specimens compared to standard mannitol-salt agar supplemented with 6 mg of oxacillin/liter.

Evaluation of an Algorithmic Approach in Comparison with the Illumigene Assay for Laboratory Diagnosis of Clostridium difficile Infection

ABSTRACT The following three diagnostic algorithms were evaluated in comparison with the Illumigene assay as a stand-alone test for Clostridium difficile detection: glutamate dehydrogenase antigen screen (GDH) followed by toxin A/B antigen testing (Tox A/B) with the cell cytotoxicity assay for discordant specimens (algorithm 1), GDH followed by the Illumigene (algorithm 2), and GDH followed by Tox A/B with the Illumigene for discordant specimens (algorithm 3). A total of 428 stool specimens submitted to three clinical microbiology laboratories in Manitoba, Canada, for C. difficile detection between June 2011 and April 2012 were included in the study. The prevalence of C. difficile in the stool specimens was 14.7% (63/428) based on toxigenic culture (microbiologic reference standard). The sensitivity and specificity of the Illumigene for C. difficile detection were 73.0% and 99.7%, respectively. The corresponding sensitivities and specificities were 65.1% and 100.0% for algorithm 1, 68.3% and 100.0% for algorithm 2, and 69.8% and 100.0% for algorithm 3. Using algorithm 1, a cell cytotoxicity assay was required for toxin detection in 37% of positive tests, prolonging turnaround time. However, the predictive value of a positive test based on a clinical reference standard (all tests positive or cytotoxigenic culture positive and clinical disease on chart review) was slightly higher with algorithm 1 than with the Illumigene assay as a stand-alone test or as part of an algorithm (algorithms 2 and 3). Based on a reduction in turnaround time, simplicity, and acceptable sensitivity and specificity, we recommend algorithm 2 (screening with the GDH antigen test and confirmatory testing with the Illumigene).

Evaluation of BacT/Alert 3D Automated Unit for Detection of Nontuberculous Mycobacteria Requiring Incubation at 30°C for Optimal Growth

ABSTRACT The reliability of the BacT/Alert 3D unit for automated detection of nontuberculous mycobacteria (NTM) that grow optimally at 30°C was assessed. This system reliably maintained a temperature of 30°C and detected 50% of the clinical NTM strains (5 Mycobacterium marinum and 3 Mycobacterium gordonae strains) faster than 37°C culture.

Continuing performance feedback and use of the ultraviolet visible marker to assess cleaning compliance in the healthcare environment

BACKGROUND Environmental surfaces have long been suspected to be a reservoir that could contribute to the presence of micro-organisms in healthcare facilities. The objective of this study was to evaluate the effect of providing weekly feedback to the housekeeping staff in improving and sustaining cleaning compliance when using ultraviolet visible marker (UVM) as an audit tool. METHODS The housekeeping staff selected 90% as the cleaning compliance target. The UVM was applied to the toilet seat, sink, soap dispenser and door knob surfaces within the patients washrooms on consecutive weekdays. The study included three arms: staff in arm 1 received cleaning compliance feedback throughout the 24-week study period, arm 2 and arm 3 staff received feedback for weeks 13-24 and weeks 1-12, respectively. Feedback was also provided to housekeeping staff by posting graphs on the wards and in the housekeeping office. FINDINGS A pre-study audit showed 66.9%, 66.5% and 66.4% cleaning compliance for arms 1, 2 and 3 respectively. While receiving weekly feedback, all three arms demonstrated significantly improved cleaning compliance (86.7%, 80.4% and 73.7% for arms 1, 2 and 3, respectively). The use of casual staff may have contributed to difficulty in achieving better cleaning compliance as arms 1, 2 and 3 had 16.1%, 26% and 40.3% of shifts filled by casual staff, respectively. CONCLUSIONS The use of UVM as an audit tool combined with weekly feedback of results to housekeeping staff resulted in significant, sustained improvement in the overall level of cleaning compliance of housekeeping staff.

CHROMagar Orientation Medium Reduces Urine Culture Workload

ABSTRACT Microbiology laboratories continually strive to streamline and improve their urine culture algorithms because of the high volumes of urine specimens they receive and the modest numbers of those specimens that are ultimately considered clinically significant. In the current study, we quantitatively measured the impact of the introduction of CHROMagar Orientation (CO) medium into routine use in two hospital laboratories and compared it to conventional culture on blood and MacConkey agars. Based on data extracted from our Laboratory Information System from 2006 to 2011, the use of CO medium resulted in a 28% reduction in workload for additional procedures such as Gram stains, subcultures, identification panels, agglutination tests, and biochemical tests. The average number of workload units (one workload unit equals 1 min of hands-on labor) per urine specimen was significantly reduced (P < 0.0001; 95% confidence interval [CI], 0.5326 to 1.047) from 2.67 in 2006 (preimplementation of CO medium) to 1.88 in 2011 (postimplementation of CO medium). We conclude that the use of CO medium streamlined the urine culture process and increased bench throughput by reducing both workload and turnaround time in our laboratories.

Evaluation of MRSASelect™ Chromogenic Medium for the Early Detection of Methicillin-Resistant Staphylococcus aureus from Blood Cultures

INTRODUCTION Staphylococcus aureus bacteremia is associated with considerable morbidity and mortality. In theory, reducing the turnaround time in reporting of methicillin-resistant S aureus (MRSA) among patients with bactermia could assist with the rapid optimization of antimicrobial therapy. OBJECTIVE To evaluate the sensitivity and specificity of MRSASelect (Bio-Rad Laboratories, USA), a chromogenic medium, in the early detection of MRSA from blood cultures growing Gram-positive cocci in clusters, and to confirm that routine use of this medium would, in fact, reduce turnaround time for MRSA identification. METHODS The present study was conducted at three microbiology laboratories in Manitoba. Between April 2010 and May 2011, positive blood cultures with Gram-positive cocci in clusters visualized on Gram stain were subcultured to both MRSASelect and routine media. MRSA isolates were identified using conventional microbiological methods from routine media and using growth with the typical colony morphology (pink colony) on MRSASelect medium. RESULTS A total of 490 blood cultures demonstrating Gram-positive cocci in clusters on Gram stain were evaluated. S aureus was recovered from 274 blood cultures, with 51 S aureus isolates (51 of 274 [18.6%]) identified as MRSA. MRSASelect medium had a sensitivity of 98%, a specificity of 100%, a positive predictive value of 100% and a negative predictive value of 99.8% for the recovery and identification of MRSA directly from positive blood culture bottles. In addition, use of MRSASelect medium was found to improve turnaround time in the detection of MRSA by almost 24 h relative to conventional methods. DISCUSSION These data support the utility of MRSASelect medium for the rapid identification of MRSA from positive blood cultures. Further clinical studies are warranted to determine whether the improvement in turnaround time will result in a measurable reduction in suboptimal antimicrobial therapy and/or improvement in patient outcome.

Ureaplasma parvum as a Cause of Sternal Wound Infection

ABSTRACT Few reports in the literature have documented the isolation of Ureaplasma species from sternal wounds. A case of sternal wound infection likely due to Ureaplasma parvum is described. When routine bacterial cultures from a sternal wound infection fail to yield a pathogen, diagnostic testing for mycoplasmas and ureaplasmas should be considered.

Re-evaluation of rejection criteria for endotracheal tube (ETT) specimens from adult patients

The purpose of this study was to determine optimal criteria for microbiology laboratory screening of endotracheal tube (ETT) specimens submitted for bacterial culture from adult patients. ETT specimens from adult patients that were received by two microbiology laboratories were prospectively evaluated and subdivided into one of three study arms with the following criteria: <10 squamous epithelial cells (SECs) per low-power field with bacteria seen on Gram staining (arm 1), >10 SECs per low-power field with bacteria seen on Gram staining (arm 2) and <10 SECs per low-power field with no bacteria seen on Gram staining (arm 3). A fourth study arm (>10 SECs per low-power field with no bacteria seen on Gram staining) was planned but this arm was terminated due to the paucity of specimens meeting these criteria. Isolate evaluation was performed using standard microbiology protocols. A limited chart review was undertaken at one of the institutions, only reviewing patients from which a potential pathogen was recovered on culture. In total, 141 ETT specimens were evaluated. A potential respiratory pathogen was recovered from 54, 37 and 10 % of specimens in study arms 1, 2, and 3, respectively (P<0.0001, comparing between arm 1 and arm 3). For the 23 patients included in the chart review from whom a potential pathogen was recovered on culture, respiratory infection was considered to be present in 50 % (6/12) of patients in arm 1, 66.6 % (6/9) of patients in arm 2 and 100 % (2/2) of patients in arm 3. Therapy was rarely altered based on culture results. In this study, the ETT specimens submitted for bacterial culture were of limited benefit to clinicians. The data presented here support the use of an absence of bacteria on Gram staining as a rejection criterion for ETT specimens. The criterion of >10 SECs per low-power field should be further evaluated in a prospective study of patients with an unequivocal clinical diagnosis of pneumonia.