[Evaluation of the performances of the iQ(®)200 ELITE automated urine microscopy analyser and comparison with manual microscopy method]. / Évaluation des performances de l'automate de cytologie urinaire Iris iQ(®)200 ELITE et comparaison avec la méthode manuelle microscopique.

Microscopy urinalysis is one of the last manual analyses in laboratories and it is difficult to control. The automation of this analysis permits our laboratory to standardise this test. We performed an evaluation of the iQ(®)200 ELITE's performances and compare the data to the manual microscopy method which is considered as "the gold standard". The repeatability achieved at different levels of leukocytes and erythrocytes showed CVs below 10% for pathological values. Inter-run repeatability is 5,57% and carry-over is negligible if we take into consideration the linearity of the formed particles. Compared to the manual method, sensitivity (Se) of the analyser for leukocytes and erythrocytes are respectively 94.85 and 100%, the negative predictive value (NPV) are 92.91 and 100%, respectively. The weaknesses of the specificity (Sp) and positive predictive value (PPV) of erythrocytes (24.22 and 41.92%) are due to the lack of sensibility of the manual method. Also, the weak PPV of yeasts, casts and crystals are due to the better detection of those particles with the iQ(®)200's image recognition system than with manual method. NPV for all the urine formed particles are excellent, they are between 98 and 100%. The iQ(®)200 ELITE permits us to standardise and control this test for a future accreditation of the laboratory. We also significantly increase the turn around time.

In vitro activity of temocillin against extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae strains isolated from urinary tract infections in France.

OBJECTIVES: Temocillin was introduced in 2015 in the French guidelines for the treatment of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae urinary tract infections. Little susceptibility data is available. We investigated the in vitro activity of temocillin against ESBL-producing Enterobacteriaceae isolated from samples of cytobacteriological examinations of urine. MATERIAL AND METHODS: Susceptibility testing was performed on 157 ESBL-producing E. coli and 95 ESBL-producing K. pneumoniae strains using the disk diffusion method. MICs of resistant strains were measured with the Etest method. RESULTS: Using current breakpoints, 71.3% of E. coli strains and 77.9% of K. pneumoniae strains were classified as susceptible. However, diameter and MIC breakpoints vary by country, and we reported discordance of clinical categorization between diameters and MIC determination for some strains. The measure of diameters was also sometimes difficult because of contaminating colonies within the inhibition zone. CONCLUSION: We highlighted difficulties related to the determination of temocillin susceptibility, such as culture of resistant colonies in the inhibition zone and discordance of clinical categorizations obtained with the disk diffusion method or the Etest method. Overall, 42% of tested Enterobacteriaceae had a diameter or MIC close to the current breakpoints; thus, it is necessary to determine the MIC for these strains before considering the clinical use of this molecule.

How to: accreditation of blood cultures' proceedings. A clinical microbiology approach for adding value to patient care.

BACKGROUND: Quality assurance and quality management are driving forces for controlling blood culture best practices but should not be disconnected from the end-point target, i.e. patient value. AIMS: This article is intended to help microbiologists implement blood culture accreditation that is actually beneficial to patient management. SOURCES: Experience from a nationwide taskforce for promoting quality assurance and competence in clinical microbiology laboratories, guidelines on blood culture. CONTENT: Experience in blood culture accreditation according to International standard ISO 15189 standards is provided in this review, with a particular focus on critical points that are specific to blood culture (e.g. excluding strain identification or antimicrobial susceptibility testing). Blood culture test method verification is based on risk analysis, and evaluation of the test method's performance is based on the literature review and suppliers' data. In addition, blood culture performance relies largely on the quality of its pre-analytical phase, and the test method should be monitored based on key performance indicators such as the volume of blood cultured, the contamination rate and time to transportation. Other critical key indicators include the rate of false-positive signals, the rate of positive blood cultures, the ecology associated with positive results, and the timely communication of the results to the ward during the post-analytical phase. Finally, a critical analysis of quality controls and of the tools needed to improve blood culture monitoring in the future is provided. IMPLICATION: Appropriate quality assurance should focus on patient value rather than technical details to provide an appropriate clinical service.

In vitro susceptibility to mecillinam of Escherichia coli strains isolated from the urine of pregnant women.

OBJECTIVES: Pivmecillinam is a safe beta-lactam for use in pregnancy. It has been widely used for the treatment of lower urinary tract infections (UTIs) in the Nordic countries where its efficacy, minor impact on the microbiota, and low level of resistance among the Escherichia coli strains have been proven. However, susceptibility data related to E. coli involved in asymptomatic bacteriuria and lower UTIs in pregnant women is lacking. We aimed to support the 2015 recommendations issued by the French Infectious Diseases Society (SPILF) on gestational UTI, with a particular focus on pivmecillinam. MATERIAL AND METHODS: Antimicrobial susceptibility testing was performed by 12 hospitals with a maternity department on 235 E. coli strains isolated from the urine of pregnant women. Susceptibility to mecillinam was tested by disk diffusion method using the 2015 recommendations of the antibiogram committee of the French microbiology society (CA-SFM). RESULTS: Global susceptibility to mecillinam was 86.4%. Susceptibility to mecillinam was 96.5% for strains susceptible to amoxicillin-clavulanic acid and 38.7% for resistant strains. All six extended-spectrum beta-lactamase-producing E. coli strains were susceptible to mecillinam. CONCLUSION: Given the efficacy and safety of pivmecillinam during pregnancy, it may be used for the documented treatment of asymptomatic bacteriuria and acute cystitis in pregnant women. It also represents an alternative for the treatment of multidrug-resistant bacterial infections.

Evaluating the management of 493 patients presenting with bacteremia in 23 northern French hospitals.

OBJECTIVES: We aimed to update the epidemiology of bacteremia and evaluate their management and short-term outcome. METHODS: We conducted a prospective multicenter survey from October to November 2011. Consecutive patients with at least one positive blood culture (BC) were included in the study. We evaluated the type and adequacy of empirical and documented antibiotic therapy, time to active antibiotic therapy, compliance with guidelines, and 10-day outcome. RESULTS: A total of 23 public and private hospitals and 633 patients (493 true pathogens and 140 contaminants) were included in the study. Patients' wards were medicine (57%), surgery (19%), intensive care (14%), onco/hematology (3.7%), pediatrics (3.4%), infectious diseases (1.8%), and obstetrics (1.2%). Main pathogens were Escherichia coli (36%), Staphylococcus aureus (16%), coagulase-negative staphylococci, and Klebsiella sp. (8% each). A total of 43 (8.7%) multidrug-resistant strains were observed, including 26 extended-spectrum beta-lactamase strains and 15 methicillin-resistant S. aureus strains. An antibiotic active against the isolated pathogen was used in 74% of empirical and 96% of documented therapies. Median time between BC and administration of an active drug was 0.61 day. Empirical antibiotic therapies were protocol-compliant in 77% of cases. Few (4%) patients with contaminated BC received an antibiotic therapy (all inappropriate). Day-10 mortality was 12.1%, higher in patients presenting with severe sepsis or septic shock (22.5%) than in patients presenting with non-severe bacteremia (7.1%; P<0.0001). CONCLUSION: The management of bacteremia seems satisfactory in these volunteer hospitals but bacteremia remains a severe infection.

Investigation of a nosocomial outbreak of extended-spectrum beta-lactamase VEB-1-producing isolates of Acinetobacter baumannii in a hospital setting.

A nosocomial outbreak of epidemiologically related VEB-1 extended-spectrum beta-lactamase-producing isolates of Acinetobacter baumannii occurred in 33 patients in an intensive care unit. A case-control study identified previous treatment with third-generation cephalosporins as the only risk factor for A. baumannii acquisition. Rationale for antibiotic use should be strengthened.

Candida albicans blastoconidia in peripheral blood smears from non-neutropenic surgical patients.

An 80 year old woman developed fever 11 days after volvulus surgery. A peripheral blood smear showed numerous yeast cells--both extraleucocytic and intraleucocytic--as well as leucoagglutination. The fungal elements included blastospores, pseudohyphae, and germ tubes. Two days later, blood cultures yielded Candida albicans, Enterobacter aerogenes, and Staphlococcus aureus. The patient had no medical history of immunodeficiency. Several reports indicate that fungal elements may be detected in peripheral blood smears from patients who have a severe intestinal disease.

Susceptibility of Enterobacteriaceae to beta-lactam agents and fluoroquinolones: a 3-year survey in France.

OBJECTIVE: To assess trends in the susceptibility to beta-lactam agents and to fluoroquinolones of clinically relevant Enterobacteriaceae isolated over a 3-year period in 14 French hospital laboratories. METHODS: During the second quarter of 1996, 1997 and 1998, 180 consecutive non-duplicate isolates of Enterobacteriaceae were collected in each center. Sixteen beta-lactams and four quinolones were tested by the disk diffusion method. In addition, the double-disk synergy test was used to screen for the production of extended-spectrum beta-lactamase (ESBL). RESULTS: Totals of 2507, 2312 and 2506 clinical isolates were obtained in each period, respectively. The distribution of Enterobacteriaceae species according to clinical specimens and wards was similar in each study period. No significant variation in the susceptibility rates to beta-lactams and fluoroquinolones was observed, except in Klebsiella pneumoniae and Enterobacter aerogenes. The prevalence of ESBL-producing isolates decreased from 18% to 9% in the former, while it increased from 32% to 54% in the latter. At the same time, the susceptibility to ofloxacin and pefloxacin increased for K. pneumoniae (P < 0.003) and cephalosporinase-producing species (P < 0.05), except Enterobacter spp. CONCLUSION: Over the 3-year study period beta-lactams and fluoroquinolones remained highly active against Enterobacteriaceae clinical isolates, with the exception of E. aerogenes, probably as a result of the dissemination of multiresistant clones in French hospitals.

Surveillance of methicillin-resistant Staphylococcus aureus (MRSA) and Enterobacteriaceae producing extended-spectrum beta-lactamase (ESBLE) in Northern France: a five-year multicentre incidence study.

In order to measure the incidence of methicillin-resistant Staphylococcus aureus (MRSA) and of Enterobacteriaceae producing extended-spectrum beta-lactamase (ESBLE), and to evaluate the impact of the national guidelines for multidrug-resistant bacteria (MDRB) prevention in hospitals of Northern France, a multicentre study was conducted for three months every year starting in 1996, in volunteer hospital laboratories. All clinical specimens positive for MRSA and ESBLE were prospectively surveyed. During the five-year surveillance period, the overall proportion of MRSA was 38.4% in the 28,534 strains of S. aureus, and that of ESBLE was 11.4% in the 6121 strains of Klebsiella pneumoniae and 47.7% in the 2353 strains of Enterobacter aerogenes. The overall incidence rates of clinical specimens positive for MRSA, ESBL-K. pneumoniae and E. aerogenes were 0.84. 0.05 and 0.12/1000 hospital-days (HD), respectively. In the 23 hospitals that participated in the survey every year, the proportion and incidence of ESBLE decreased. Hence, despite recommendations as for isolation precautions, MRSA remains poorly controlled and requires more effective measures.

Relapsing Plasmodium vivax malaria with atypical parasite forms and phagocytosis by peripheral neutrophils.

A case of atypical Plasmodium vivax malaria is presented. The clinical follow-up has allowed to characterize three consecutive malaria clinical episodes within one year. At the first attack, 39% of the infected red blood cells were parasitized by gametocytes. Furthermore, rare crisis forms, exceptional "pseudoparthenogenesis" forms, a few equatorial trophozoites, malaria pigment-containing leucocytes and phagocytized parasites were also found in the thin blood smears. At the second malaria episode, morphological aspects were quite similar, but the gametocyte percentage decreased and that of the equatorial trophozoite forms increased. Only at the third attack, was the morphology typical of P. vivax. The Plasmodium species and the absence of mixed infection were unequivocally confirmed using polymerase chain reaction. Atypical strains of P. vivax are relatively frequent. Nevertheless, to our knowledge, neither so high a gametocyte percentage, nor extensive P. vivax peripheral phagocytosis were previously reported.

[Evaluation of a dot-blot technique for the detection of Epstein-Barr virus antibodies (IgG anti-viral capsid antigen)]. / Evaluation d'une technique de dot-blot pour la détection d'anticorps dirigés contre le virus d'Epstein-Barr (IgG anti-antigène de capside virale).

Infectious mononucleosis syndrome is caused by several infectious agents, including Epstein-Barr virus (EBV), cytomegalovirus (CMV), and Toxoplasma gondii. The ImmunoDot EBV VCA-IgG test (Biomedical Diagnostics) is a dot-blot enzyme-immunoassay, which determines the presence of heterophile antibodies and IgG antibodies directed against Epstein-Barr viral capsid antigen (anti-VCA), CMV and T. gondii. This test is simple, unitary, ready-to-use, and rapid (40 minutes), requiring approximately 10 microliters of serum or plasma or 20 microliters of whole blood. The aim of this study was to compare the results of this technique for the determination of anti-VCA IgG antibodies with those obtained by the reference technique using indirect immunofluorescence on 185 serum specimens. A sensitivity of 99.2%, a specificity of 92.4%, a correlation with indirect immunofluorescence of 97.3%, a the absence of cross-reactions with CMV, T. gondii, and herpes simplex viruses and the absence of interference with antinuclear antibodies were the main results of this comparative study. In association with another test (Monolert 2TM) detecting IgM and IgG antibodies against Epstein-Barr nuclear antigen, the dot-blot method represents a useful screening strategy for EBV response.

[Clostridium difficile bacteremia]. / Bactériémie à Clostridium difficile.

BACKGROUND: Extra-digestive manifestations of Clostridium difficile infection are very uncommon. Exceptional cases of C. difficile bacteremia or severe sepsis have been described in intensive care patients, demonstrating the capacity of this agent to generate generalized infection. CASE REPORT: C. difficile bacteremia occurred in a 66-year-old immunodepressed patient treated for acute myeloblastic leukemia. Bacteremia was associated with a abscess of the anal margin. Outcome was favorable after treatment with metronidosole. DISCUSSION: Clostridium difficile is generally selected by prior antibiotic treatment. It is the principal agent of nosocomial diarrhea. In immunodepressed patients, systemic dissemination is a rare but possible development.

[Lung infection by Mycobacterium szulgai]. / Infection pulmonaire à Mycobacterium szulgai.

BACKGROUND: Mycobacterium szulgai is an uncommon atypical mycobacterium human pathogen. CASE REPORT: The clinical manifestations and radiographic findings in a 31-year-old woman were strongly suggestive of pulmonary tuberculosis. The mode of transmission could not be determined. Mycobacterium szulgai was identified. The patient was treated with antituberculosis antibiotics and the clinical course was favorable. DISCUSSION: Mycobacterium szulgai is an atypical mycobacterrium difficult to identify. Its epidemiological features are unknown. This potential pulmonary pathogen is rarely described in the literature. Most cases have involved pulmonary, bone and joint or skin infections in immunodepressed patients. M. szulgai is relatively susceptible to classic antituberculosis antibiotics although standard regimens have not been established. Our patient required intensive care for mechanical ventilation.

Evaluation of glycopeptide use in nine French hospitals.

BACKGROUND: Adequate glycopeptide use is necessary to optimize efficacy and minimize adverse events and selection of drug resistant microorganisms. METHODS: We performed a multicenter prospective observational study of glycopeptides indications based on modified HICPAC criteria, and quality of prescription based on French consensus recommendations. We assessed the adequacy of indications, loading dose, maintenance dose, administration frequency, serum monitoring and its consequences. RESULTS: We evaluated 117 curative adult prescriptions from nine hospitals. Glycopeptide indications were adequate in 71% with either a clinical (55%) or microbiological justification (27%). The indication was more frequently adequate for vancomycin than for teicoplanin (77% vs. 60%, p=0.04). De-escalation therapy was performed in 48% of all documented infections and 78% of documented infections with betalactam susceptible strains. We observed high rates of correct maintenance dose (90%), administration frequency (88%), and serum monitoring (74%). Loading doses (43%), adequate serum monitoring timing (43%), correction of dose because of inadequate levels (32%) were less adequate. Overall, only three (3%) treatments were adequate for all evaluated items. CONCLUSION: Glycopeptide indications were adequate. However, the quality of prescription can be improved and should focus on loading doses and serum monitoring.

[Quantitative and qualitative study of the fecal flora of cirrhotic patients]. / Etude quantitative et qualitative de la flore fécale du cirrhotique.

In this study, the qualitative and quantitative fecal flora was determined in 30 cirrhotic patients. The flora of these patients was composed of some bacterial populations and does not differ appreciably from the normal.

[Value of spiral automatic plating in quantitative bacteriology]. / Intérêt de l'ensemenceur automatique spiral en bactériologie quantitative.

The Spiral plate count method, semi-automated plating technique, presents some advantages: saving of time and materials, fast and easy counting. This method appears well adapted for studies of microbial ecology.