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Microbiology Laboratory (microbiology + laboratory)
Selected AbstractsPATHOGEN DETECTION IN FOOD MICROBIOLOGY LABORATORIES: AN ANALYSIS OF QUALITATIVE PROFICIENCY TEST DATA, 1999,2007JOURNAL OF FOOD SAFETY, Issue 4 2009DANIEL C. EDSON ABSTRACT The objective of this study was to assess laboratories' ability to detect or rule out the presence of four common food pathogens: Escherichia coli O157:H7, Salmonella spp., Listeria monocytogenes and Campylobacter spp. To do this, qualitative proficiency test data provided by one proficiency test provider from 1999 to 2007 were examined. The annual and cumulative 9-year percentages of false-negative and false-positive responses were calculated. The cumulative 9-year false-negative rates were 7.8% for E. coli O157:H7, 5.9% for Salmonella spp., 7.2% for L. monocytogenes and 13.6% for Campylobacter spp. Atypical strains and low concentrations of bacteria were more likely to be missed, and the data showed no trend of improving performance over time. Percentages of false-positive results were below 5.0% for all four pathogens. PRACTICAL APPLICATIONS The results imply that food testing laboratories often fail to detect the presence of these four food pathogens in real food specimens. To improve pathogen detection, supervisors should ensure that testing personnel are adequately trained, that recommended procedures are followed correctly, that samples are properly prepared, that proper conditions (temperature, atmosphere and incubation time) are maintained for good bacterial growth and that recommended quality control procedures are followed. Supervisors should also always investigate reasons for unsatisfactory proficiency test results and take corrective action. Finally, more research is needed into testing practices and proficiency test performance in food testing laboratories. [source] Comparison of the performance of rapid HIV tests using samples collected for surveillance in Mozambique,JOURNAL OF MEDICAL VIROLOGY, Issue 12 2009Josefa Melo Abstract Mozambique had low HIV prevalence until the mid-1990s, but recent data indicate increasing rates. There is little information on HIV-2. Therefore, HIV seroprevalence was assessed among pregnant women and field-ready HIV diagnostic strategies were evaluated. A total of 6,930 samples collected by three health centers from 2002 to 2005 were tested on site by nurses with two simple/rapid tests, Determine HIV-1/2 (Abbott Laboratories; screening) and Uni-Gold HIV (Trinity Biotech; confirmation), which is the national HIV testing strategy. The prevalence of HIV was 14.0% (2002), 17.8% (2003), 16.5% (2004), and 20.2% (2005). A subset of 888 samples collected 2003 was sent to the Central Microbiology Laboratory, Maputo for evaluation of tests and testing strategies. The assays included for comparison were Capillus HIV-1/HIV-2 (Trinity Biotech), DoubleCheckGold HIV-1&2 (Orgenics) and Enzygnost Anti-HIV-1/2 Plus (Behringwerke, reference ELISA). Confirmation of reactive samples was done by Uni-Gold HIV and ImmunoComb II HIV-1&2 BiSpot (for HIV type differentiation). The Capillus HIV-1/ HIV-2,+,ImmunoComb II HIV-1&2 BiSpot combination was the gold standard. The sensitivity of the rapid/simple screening assays (Determine HIV-1/2, DoubleCheckGold HIV-1&2) was 100% (N,=,160) and their (initial) specificities were 99.6% and 99.7%, respectively. Repeated testing and combinations of assays increased the specificity. Four suspected cases of recent seroconversion were found. Together with the increasing prevalence rates, this may indicate that Mozambique is a high-incidence area, although further studies are needed to confirm this. Testing strategies for on-site screening and confirmation based on the combination of Determine HIV-1/2, Uni-Gold HIV and DoubleCheckGold HIV-1&2 are well suited for local field use. J. Med. Virol. 81:1991,1998, 2009. © 2009 Wiley-Liss, Inc. [source] Bacterial dormancy in Campylobacter: abstract theory or cause for concern?INTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 6 2001John E. Moore For the past 100 years, since the birth of modern microbiology, this discipline has predominantly relied on the ability to culture micro-organisms in vitro on artificial synthetic culture media under controlled conditions in the laboratory. However, sometimes it is not possible to detect foodborne pathogens using such conventional techniques. Employment of these techniques can also lead to a delay in detection of pathogens. The ,viable but non-culturable' (VNC) cellular form has been demonstrated in Campylobacter jejuni, representing a resting or dormant stage, which is induced through cell stress including starvation. This form is extremely difficult to detect and generally requires complex and sophisticated technology which is usually not available in most routine food microbiology laboratories. This review aims at examining the role of this cell form in Campylobacter, including their historical evolution, formation, physiology, detection and to discuss the challenges that this form presents to food safety. [source] Wide variation in effectiveness of laboratory disinfectants against bacteriophagesLETTERS IN APPLIED MICROBIOLOGY, Issue 6 2008D.E. Halfhide Abstract Aims:, The purpose of this study was to identify an effective disinfectant for the inactivation of the bacteriophages (phages) being used in our laboratory, as published studies on phage inactivation are far from unanimous in their conclusions. Methods and Results:, The phages studied were three closely related strains of Myoviridae and three strains of Siphoviridae. Three disinfectants which are used commonly in microbiology laboratories were evaluated: Virkon (1%), ethanol (75%) and sodium hypochlorite (2500 ppm available chlorine). The most effective of these was Virkon, which inactivated all six phages rapidly. Ethanol was effective against the Myoviridae but had little effect on the Siphoviridae. Sodium hypochlorite was the least effective of the disinfectants evaluated. Conclusions:, The findings of this study demonstrate a wide diversity in the effectiveness of disinfectants tested for inactivation of phages. Significance and Impact of the Study:, Of the disinfectants tested Virkon is the most suitable choice for those unable to carry out disinfection validation studies, or where a broad spectrum disinfectant against phages is required. All of the phages in this study showed resilience to inactivation by sodium hypochlorite, and therefore this disinfectant is an unwise choice for use against phage without first assessing its effectiveness. [source] Feasible identification of Staphylococcus,epidermidis using desferrioxamine and fosfomycin disks,APMIS, Issue 1 2008ANA LÚCIA SOUZA ANTUNES Coagulase-negative Staphylococcus spp. (CoNS) have emerged as predominant pathogens in hospital-acquired infections, as well as reservoirs of antimicrobial resistance, increasing the necessity of developing reliable methods for identification of the most frequent species. The aim of this study was to propose a simplified method for identification of Staphylococcus epidermidis. A total of 490 isolates of CoNS were identified by Bannerman's method. Taking into account distinct approaches for identification of S. epidermidis, among CoNS, we proposed the use of only two disks: desferrioxamine for the initial trial, and fosfomycin to match the final identification. Of the 320 isolates susceptible to desferrioxamine, Bannerman's method identified 238 S. epidermidis and 73 S. hominis, while we achieved identification of 239 S. epidermidis and 76 S. hominis. Compared to Bannerman's method, the method proposed here obtained a sensitivity of 99.5%, and had a positive predictor value of 99.2%. We also used a genotypic method for identification of S. epidermidis by polymerase chain reaction (PCR) targeting the tuf gene. In conclusion, the method proposed here has proved to be useful for the identification of S. epidermidis, the most frequent species of CoNS isolated from blood cultures in clinical microbiology laboratories. [source] High-throughput epidemiologic typing in clinical microbiologyCLINICAL MICROBIOLOGY AND INFECTION, Issue 2 2003A. Van Belkum Mapping, and ultimately preventing, the dissemination of infectious agents is an important topic in public health. Newly developed molecular,microbiological methods have contributed significantly to recent advances in the efficient tracking of the nosocomial and environmental spread of microbial pathogens. Not only has the application of novel technologies led to improved understanding of microbial epidemiology, but the concepts of population structure and dynamics of many of the medically significant microorganisms have advanced significantly also. Currently, genetic identification of microbes is also within the reach of clinical microbiology laboratory professionals including those without specialized technology research interests. This review summarizes the possibilities for high-throughput molecular,microbiological typing in adequately equipped medical microbiology laboratories from both clinical and fundamental research perspectives. First, the development and application of methods for large-scale comparative typing of serially isolated microbial strains are discussed. The outcome of studies employing these methods allows for long-term epidemiologic surveillance of infectious diseases. Second, recent methods enable an almost nucleotide-by-nucleotide genetic comparison of smaller numbers of strains, thereby facilitating the identification of the genetic basis of, for instance, medically relevant microbiological traits. Whereas the first approach provides insights into the dynamic spread of infectious agents, the second provides insights into intragenomic dynamics and genetic functionality. The current state of technology is summarized, and future perspectives are sketched. [source] Etiologic spectrum and pattern of antimicrobial drug susceptibility in bacterial meningitis in Sokoto, NigeriaACTA PAEDIATRICA, Issue 8 2000FE EmeleArticle first published online: 2 JAN 200 Etiologic agents of meningitis were prospectively investigated among patients admitted to Usman Danfodio University Teaching Hospital, Sokoto. Of 1097 cerebrospinal fluid (CSF) samples submitted to the microbiology laboratory from various wards of the hospital, 289 (26%) were microscopically, culturally and/or serologically proven to be bacterial meningitis. The etiologic spectrum was as follows: Neisseria meningitidis (61%), Streptococcus pneumoniae (18%), Haemophilus influenzae (10%), Staphylococcus aureus (6%), Coliform bacilli (3%), Escherichia coli (0.7%), Mycobacterium tuberculosis (0.7%), Listeria monocytogenes (0.4%), Flavobacterium meningosepticum (0.4%) and Pseudomonas putrifasciens (0.4%). Bacterial meningitis was most prevalent (195 or 68%) among children aged 1-9 y, while adults and neonates were least affected. Coliform bacilli caused five of eight neonatal cases. Males were more frequently affected than females (x2=12.50;p < 0.05). Culture and microscopy were comparatively less efficient than the search for bacterial antigens, especially in the diagnosis of Haemophilus meningitis. Antimicrobial susceptibility of N. meningitidis to ampicillin and benzyl penicillin reduced progressively over the years (F = 406.98;p < 0.001). Nineteen (11%) of the isolates (5 Meningococci, 7 Staph. aureus, 1 Haem. influenza and 6 others) showed simultaneous resistance to chloramphenicol, ampicillin and benzyl penicillin. [source] Mailed urine samples are not an effective screening approach for Chlamydia trachomatis case finding among young menJOURNAL OF THE EUROPEAN ACADEMY OF DERMATOLOGY & VENEREOLOGY, Issue 6 2007M Domeika Abstract Background, Frequency of testing is known to be low for sexually transmitted infections (STIs) in men aged 20,24 years. The use of mailed, home-obtained urine specimens could increase the uptake of young men and facilitate screening programmes for the detection of asymptomatic Chlamydia trachomatis. Objective, The aim of the present study is to evaluate the home screening approach as a tool for recruitment of asymptomatic men for screening of genital C. trachomatis infections. Methods, Men aged 19,24 years old (n = 1936) were invited to participate in home-based testing for genital C. trachomatis infection. Persons who agreed to be tested were provided with a testing kit. Self-collected first void urine was sent for testing to the microbiology laboratory. The test result was accessible on the study's web-page 1 week after testing. Individuals with a diagnosed infection were instructed to contact the venereal disease department. Results, The response rate was 24% (462/1936). The responders' main reason for not participating was a feeling of being safe regarding STIs (87%; 159/182). The primary reason for this feeling of safety was that the responders were in a steady relationship (59%; 107/159). Having sex outside a steady relationship was reported by 36% (90/250) of the responders. The prevalence of C. trachomatis infection among the responders was 2.02% and the reported history of chlamydial infection was 36% (34/95). Out of the responders, 92% (229/249) were, to varying degrees, concerned about getting STIs; however, the majority (72%; 174/242) estimated the risk to be low. Conclusion, Home screening using web-based answer management is a feasible tool for STI screening, which lowers the threshold for people at risk. In this particular population, however, the response rate was too low to be routinely introduced. [source] Performance of commercial latex agglutination tests for the differentiation of Candida dubliniensis and Candida albicans in routine diagnostics,APMIS, Issue 11 2007E. CHRYSSANTHOU Candida dubliniensis is phenotypically similar to Candida albicans and may therefore be underdiagnosed in the clinical microbiology laboratory. The performance of Bichro-Dubli latex agglutination test for rapid species identification of C. dubliniensis was prospectively evaluated on 111 vaginal and 118 respiratory isolates. These had presumptively been identified as C. albicans/C. dubliniensis by their green colonies on CHROMagar Candida plates. Bichro-Dubli test identifed 2 (1.8%) vaginal and 6 (5.1%) respiratory isolates as C. dubliniensis. The test was also positive for 37 C. dubliniensis control strains characterised by 18S-28S DNA-sequencing. Bichro-Dubli test is thus a sensitive and accurate tool for rapid diagnostics in routine laboratories. [source] Use of Quantitative Broad-based Polymerase Chain Reaction for Detection and Identification of Common Bacterial Pathogens in Cerebrospinal FluidACADEMIC EMERGENCY MEDICINE, Issue 7 2010Richard Rothman MD ACADEMIC EMERGENCY MEDICINE 2010; 17:741,747 © 2010 by the Society for Academic Emergency Medicine Abstract Background:, Conventional laboratory diagnosis of bacterial meningitis based on microscopy followed by culture is time-consuming and has only moderate sensitivity. Objectives:, The objective was to define the limit of detection (LOD), analytic specificity, and performance characteristics of a broad-based quantitative multiprobe polymerase chain reaction (PCR) assay for rapid bacterial detection and simultaneous pathogen-specific identification in patients with suspected meningitis. Methods:, A PCR algorithm consisting of initial broad-based detection of Eubacteriales by a universal probe, followed by pathogen identification using either pathogen-specific probes or Gram-typing probes, was employed to detect pathogens. The 16S rRNA gene, which contains both conserved and variable regions, was chosen as the target. Pathogen-specific probes were designed for Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenzae, Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli, and Listeria monocytogenes. Gram-positive and -negative typing probes were designed based on conserved regions across all eubacteria. The LOD and time to detection were assessed by dilutional mocked-up samples. A total of 108 convenience cerebrospinal fluid (CSF) clinical samples obtained from the Johns Hopkins Hospital (JHH) microbiology laboratory were tested, and results were compared with hospital microbiologic culture reports. Results:, The LOD of the assay ranged from 101 to 102 colony-forming units (CFU)/mL. Pathogen-specific probes showed no cross-reactivity with other organisms. Time to detection was 3 hours. In clinical specimens, the universal probe correctly detected 16 of 22 culture-positive clinical specimens (sensitivity = 72.7%; 95% confidence interval [CI] = 49.8% to 89.3%), which were all correctly characterized by either pathogen-specific or Gram-typing probes. Adjusted sensitivity after removing probable microbiologic laboratory contaminants was 88.9% (95% CI = 65.3% to 98.6%). The universal probe was negative for 86 of 86 culture-negative specimens. Conclusions:, A broad-based multiprobe PCR assay demonstrated strong analytic performance characteristics. Findings from a pilot clinical study showed promise in translation to human subjects, supporting potential utility of the assay as an adjunct to traditional diagnostics for early identification of bacterial meningitis. [source] Performance of two tube coagulase methods for rapid identification of Staphylococcus aureus from blood cultures and their impact on antimicrobial managementCLINICAL MICROBIOLOGY AND INFECTION, Issue 5 2008P. D. J. Sturm Abstract Test parameters and clinical impact of the direct tube coagulase test (DTCT) for rapid identification of Staphylococcus aureus from blood culture were investigated. The sensitivity of the DTCT at 4 h using saline dilution was 96%, compared with 93% using serum separator tubes; specificity was 100% for both methods. Among 32 patients with S. aureus bacteraemia, treatment modifications were based on microbiology results from the primary source of infection in 12 patients, on a Gram's stain from blood culture in seven patients, and on the DTCT in nine patients. The DTCT is a valuable adjunct in the routine microbiology laboratory because of its good performance, technical simplicity and low cost. [source] | ||||||||||||||||