Development and evaluation of a multiplex test for the detection of atypical bacterial DNA in community-acquired pneumonia during childhood

An incorrect or late diagnosis can lead to an increase in the morbidity and mortality caused by pneumonia, and the availability of a rapid and accurate microbiological test to verify the aetiology is imperative. This study evaluated a molecular test for the identification of the bacterial cause of atypical community-acquired pneumonia (ACAP). Fifty-four children with pneumonia were studied using bacteriological cultures, Mycoplasma pneumoniae, Coxiella burnetii, Chlamydophila pneumoniae and Legionella spp. serology, and Streptococcus pneumoniae and Legionella antigens. Simultaneously, the presence of bacterial and fungal DNA was tested for in respiratory secretion samples using the Vircell SL kit, including multiplex PCR and amplicon detection by means of line blots. There were 14 cases of ACAP caused by M. pneumoniae, with positive kit results for 13 of them, and two cases of Q-fever, with negative kit results for Coxiella burnetii. The test was negative in the remaining 38 cases (one staphylococcal pneumonia, 20 Streptococcus pneumoniae pneumonias, and 17 probable viral pneumonias). The sensitivity of the test for the detection of M. pneumoniae was 92.8% and the specificity was 100%. The Vircell SL kit allows detection of M. pneumoniae DNA in respiratory secretion samples from children with ACAP

Different presence of Chlamydia pneumoniae, herpes simplex virus type 1, human herpes virus 6, and Toxoplasma gondii in schizophrenia: meta-analysis and analytical study

In the present study we have performed both a meta-analysis and an analytical study exploring the presence of Chlamydia pneumoniae, herpes simplex virus type 1, human herpes virus 6, and Toxoplasma gondii antibodies in a sample of 143 schizophrenic patients and 143 control subjects. The meta-analysis was performed on papers published up to April 2014. The presence of serum immunoglobulin G and immunoglobulin A was performed by enzyme-linked immunosorbent assay test. The detection of microbial DNA in total peripheral blood was performed by nested polymerase chain reaction. The meta-analysis showed that: 1) C. pneumoniae DNA in blood and brain are more common in schizophrenic patients; 2) there is association with parasitism by T. gondii, despite the existence of publication bias; and 3) herpes viruses were not more common in schizophrenic patients. In our sample only anti-Toxoplasma immunoglobulin G was more prevalent and may be a risk factor related to schizophrenia, with potential value for prevention.Part of this work was presented at the Royal Academy of Medicine of Spain

Oral fosfomycin for treatment of urinary tract infection: a retrospective cohort study

BACKGROUND: Fosfomycin is increasingly called upon for the treatment of multi drug-resistant (MDR) organisms causing urinary tract infection (UTI). We reviewed oral fosfomycin use for UTI treatment in a large UK hospital. The primary goal was to audit our clinical practice against current national guidelines. Secondary aims were to identify factors associated with treatment failure, and to investigate the potential for using fosfomycin in patients with co-morbidities. METHODS: We retrospectively studied 75 adult patients with UTI who received 151 episodes of treatment with fosfomycin from March 2013 to June 2015. We collected clinical data from our electronic patient record, and microbiology data pre- and post- fosfomycin treatment. We recorded additional data for patients receiving prolonged courses in order to make a preliminary assessment of safety and efficacy. We also reviewed >18,000 urinary tract isolates of Escherichia coli and Klebsiella spp. processed by our laboratory over the final year of our study period to determine the prevalence of fosfomycin resistance. RESULTS: There was a significant increase in fosfomycin treatment episodes over the course of the study period. Co-morbidities were present in 71 % of patients. The majority had E. coli infection (69 %), of which 59 % were extended spectrum beta-lactamase (ESBL)-producers. Klebsiella infections were more likely than E. coli to fail treatment, and more likely to be reported as fosfomycin resistant in cases of relapse following treatment. There were no adverse events in five patients treated with prolonged fosfomycin. Among all urinary isolates collected over a year, fosfomycin resistance was documented in 1 % of E. coli vs. 19 % of Klebsiella spp. (p < 0.0001). CONCLUSIONS: We report an important role for oral fosfomycin for MDR UTI treatment in a UK hospital population, and based on the findings from this study, we present our own local guidelines for its use. We present preliminary data suggesting that fosfomycin is safe and effective for use in patients with complex comorbidities and over prolonged time periods, but may be less effective against Klebsiella than E. coli

Caracterización de Enterococcus faecium no sensible a daptomicina produciendo infección del tracto urinario en un paciente trasplantado renal

This work was supported only by the National Institute of Allergy and Infectious Diseases (National Institutes of Health [NIH] grant R01 AI093749 to C. A. A.)Objectives: Characterization of a urine isolate of daptomycin non-susceptible Enterococcus faecium recovered from a patient with kidney transplantation and no history of daptomycin exposure. Methods: After isolation in a urine sample, identification of E. faecium was confirmed by amplification of the E. faecium-specific gene encoding D-alanyl-D-alanine ligase (ddl) and daptomycin susceptibility testing was performed by E-test on cation-adjusted Mueller-Hinton agar. In order to determine the genetic bases of daptomycin resistance, the open reading frames of five genes previously associated with daptomycin resistance in enterococci were sequenced. Results: Substitutions in the response regulator LiaR (S19F) and cardiolipin synthase (R218Q) were identified. Conclusions: To the best of our knowledge, this is the first characterization of emerging daptomycin resistance in E. faecium in a Spanish hospital in the absence of daptomycin exposure and in a renal transplant recipient.Objetivos. Presentamos la caracterización de un aislado de Enterococcus faecium no sensible a daptomicina, recuperado de una muestra de orina de un paciente con trasplante de riñón e infección urinaria y sin antecedentes de exposición previa a daptomicina. Métodos. Tras el aislamiento, la identificación de E. faecium fue confirmada por la amplificación del gen que codifica la región específica de la ligasa de la D-alanil-D-alanina (ddl) y la prueba de sensibilidad a daptomicina se realizó mediante E-test en agar Mueller-Hinton ajustado para cationes. Con el fin de determinar las bases genéticas de la resistencia a daptomicina, se secuenciaron las regiones de lectura abierta de cinco genes previamente asociados con la resistencia a daptomicina en enterococos. Resultados. Se identificaron cambios en el promotor de LiaR (S19F) y la sintetasa de la cardiolipina (R218Q). Conclusiones. Esta es la primera caracterización de un aislado clínico de E. faecium con resistencia a daptomicina en un hospital español, en ausencia de exposición previa y en un receptor de trasplante renal.National Institutes of Health [NIH] R01 AI09374

Detection of new mutations conferring resistance to linezolid in glycopeptide-intermediate susceptibility Staphylococcus hominis subspecies hominis circulating in an intensive care unit

Glycopeptides and linezolid are the most widely used antibiotics to treat infections by methicillin-resistant Staphylococcus spp. We report the presence of various isolates of methicillin-resistant S. hominis subsp. hominis with resistance to linezolid and reduced susceptibility to glycopeptides. We studied ten blood culture isolates of S. hominis subsp. hominis from nine patients admitted to our hospital. Etest was used to study susceptibility to antibiotics commonly prescribed against staphylococci. Domain V region of the 23S rRNA gene was amplified and sequenced to detect possible mutations that confer resistance to linezolid. Pulsed-field gel electrophoresis (PFGE) was used for the clonality study of isolates. All isolates were resistant to oxacillin, gentamicin, levofloxacin, cotrimoxazole, and linezolid, and susceptible to tigecycline and daptomycin. Nine of the isolates were resistant to erythromycin and clindamycin, and showed heterogeneous resistance to glycopeptides. C2190T, G2603T, and G2474T mutations were detected in domain V of the 23S rRNA gene. PFGE showed the presence of two different clones. This report alerts to the possible appearance of clinical strains of methicillin-resistant staphylococci with intermediate resistance to glycopeptides, resistance to linezolid, and multiple resistance to other second-line antibiotics