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false positive Legionella pcr

F Carinci, L Scapoli, M Contaldo, R Santoro, A Palmieri, F Pezzetti, D Lauritano, V Candotto, D Mucchi, L Baggi, A Tagliabue, L Tettamanti
Legionella spp. are ubiquitous in aquatic habitats and water distribution systems, including dental unit waterlines. Surveys have shown that the percentage of samples taken at different dental sites that were positive for Legionella spp. were highly variable and ranged from 0% to 100%. Cultivation is the principal approach to evaluating bacterial contamination employed in the past, but applying this approach to testing for Legionella spp. may result in false-negative data or underestimated bacterial counts...
January 2018: Journal of Biological Regulators and Homeostatic Agents
Tomer Avni, Amir Bieber, Hefziba Green, Tali Steinmetz, Leonard Leibovici, Mical Paul
The diagnosis of Legionnaires' disease (LD) is based on the isolation of Legionella spp., a 4-fold rise in antibodies, a positive urinary antigen (UA), or direct immunofluorescence tests. PCR is not accepted as a diagnostic tool for LD. This systematic review assesses the diagnostic accuracy of PCR in various clinical samples with a direct comparison versus UA. We included prospective or retrospective cohort and case-control studies. Studies were included if they used the Centers for Disease Control and Prevention consensus definition criteria of LD or a similar one, assessed only patients with clinical pneumonia, and reported data for all true-positive, false-positive, true-negative, and false-negative results...
February 2016: Journal of Clinical Microbiology
Katarzyna Pancer, Karina Jahnz-Różyk, Aleksandra Kucharczyk, Włodzimierz Gut, Piotr Kucharczyk, Pernille L Elverdal, Bogumiła Litwińska
UNLABELLED: The aim of this study was elimination of false positive results obtained by the Chlamylege kit. Two serological kits (IgM ELISA L. pneumophila sgs1-7; ImmuView(TM) L. pneumophila sg1/sg3) and pre-absorption tests (with L. pneumophila sg1 and sg3 reference strains antigens) were used. 153 sera (79 patients) were examined. The high correlations were found between the results by both tests. Positive results by ELISA (sgs1-7) were found in 19/79 patients; by ImmuView(TM) (sg1+sg3) in 16/63...
2012: Annals of Agricultural and Environmental Medicine: AAEM
Robert Schofield, Roger J Trent
A 59-year-old man presented with a severe flu-like illness and widespread pulmonary infiltrates on chest x-ray. A rapid influenza direct test was positive and the patient was nursed in isolation. On subsequent review, a diagnosis of probable atypical pneumonia was made, which was confirmed with positive urinary serology for Legionella pneumophila and treatment with appropriate antibiotics was started. A real-time PCR test for influenza A and B was negative at 72 h. The patient made a slow but full recovery and was discharged after 14 days...
August 13, 2010: BMJ Case Reports
William F McCoy, Erin L Downes, Lesley F Leonidas, Melissa F Cain, Daniel L Sherman, Kevin Chen, Sangeetha Devender, Michael J Neville
Most Legionella culture tests are performed on building water samples that have been shipped to analytical laboratories for analysis. Significant (≥ 1 log₁₀ unit) changes in results were observed in 52% of held samples (6 h or longer, ambient temperature) drawn from building water systems in a 42-sample initial survey. It was not practical to use the spread plate protocol for on-site "t = 0" cultures in a larger, more diverse survey of thousands of building water systems. Two thousand four hundred twenty-one (2421) building water samples were split for on-site analysis using a field culture protocol and then also cultured after overnight shipment to the lab for analysis with the standardized spread plate method...
July 2012: Water Research
Louise H Krøjgaard, Karen A Krogfelt, Hans-Jørgen Albrechtsen, Søren A Uldum
BACKGROUND: Culture and quantitative polymerase chain reaction (qPCR) assays for the detection of Legionella were compared on samples from a residential area before and after two interventions. A total of 84 samples were collected from shower hoses and taps as first flush samples and at constant temperature. Samples were grouped according to the origin of the sample, a) circulation water b) water from empty apartments c) water from shower hoses. The aims were to investigate the usefulness of qPCR compared to culture for monitoring remedial actions for elimination of Legionella bacteria and as a tool for risk assessment...
2011: BMC Microbiology
Pernille L Elverdal, Christina W Svarrer, Charlotte S Jørgensen, Ian C Skovsted, Søren A Uldum
The aim of this study was to determine whether separate measurement of immunoglobulin (Ig) M and G antibodies to Legionella (L.) pneumophila serogroups (sg) 1, 3 and 6 as single antigens can facilitate an early diagnosis of Legionnaires' disease. The developed ELISA was evaluated and compared with an in-house indirect Legionella immunofluorescence antibody test (IFAT) measuring Total Ig. A total of 193 sera from 128 patients with confirmed L. pneumophila infections were used to assess the sensitivity of the developed ELISA...
September 2011: Journal of Microbiological Methods
Iu S Aliapkina, Iu E Dronina, T I Karpova, Iu P Pashko, D A Varlamov, Ia I Alekseev, Iu M Romanova, I S Tartakovskiĭ
AIM: To assess efficacy of using the method of quantitative detection of Legionella in objects of the environment by real-time polymerase chain reaction (RT-PCR). MATERIALS AND METHODS: For the development of the assay, genus-specific primers from gene coding 16S rRNAas well as species-specific primers for detection of Legionella pneumophila on the basis of mip gene sequence. For quantitative detection of L. pneumophila calibration samples of pGEM plasmid containing fragment of the mip gene in known concentration were used...
March 2009: Zhurnal Mikrobiologii, Epidemiologii, i Immunobiologii
D O Mikhaĭlova, Z D Bobyleva, V V Bazarnyi, E P Amon, Ia B Beĭkin, L G Besedina, O V Mel'nikova, V P Shilova, S M Rozanova, E Iu Perevalova
The aim of the study was a comparative analysis of diagnostic value of different laboratoty methods conducted on the basis of results of examination of patients during Legionnaires' disease outbreak in town Verkhnyaya Pyshma. Retrospective analysis of laboratory data from 74 patients with diagnosis of Legionnaires' disease was performed. Complex of laboratory methods was used (polymerase chain reaction (PCR), enzyme immunoassay (EIA), immunochromatography). In group of patients with Legionnaires' disease, the highest proportion of positive results (73%) was obtained by the EIA determining total specific antibodies in urine...
March 2008: Zhurnal Mikrobiologii, Epidemiologii, i Immunobiologii
P Elverdal, C S Jørgensen, S A Uldum
Four commercially available kits from (1) Focus Diagnostics, (2) SERION, (3) Zeus and (4) Vircell for detection of antibodies to Legionella pneumophila were evaluated with panels of sera from patients with proven Legionella infection (n = 81) and/or other bacterial infections (n = 75). An in-house indirect Legionella immunofluorescence antibody test (IF test) was used as reference. All sera from the laboratory-proven Legionella pneumophila cases [culture, urinary antigen test and/or polymerase chain reaction] of Legionella infection were found to be positive by the in-house IF test...
February 2008: European Journal of Clinical Microbiology & Infectious Diseases
C Ginevra, C Barranger, A Ros, O Mory, J-L Stephan, F Freymuth, M Joannès, B Pozzetto, F Grattard
This study describes the development and evaluation of a new commercial test, Chlamylege (Argene Inc.), which allows the simultaneous detection in respiratory samples of Chlamydophila pneumoniae, Mycoplasma pneumoniae, and most Legionella species, as well as PCR inhibitors, by using a multiplex PCR and microplate hybridization. The sensitivities of Chlamylege were 1 x 10(-3) IFU, 5 x 10(-2) color-changing units, and 1 CFU per reaction tube for C. pneumoniae, M. pneumoniae, and Legionella pneumophila, respectively...
July 2005: Journal of Clinical Microbiology
S C Weir, S H Fischer, F Stock, V J Gill
Using polymerase chain reaction (PCR) for the detection of pathogens that are difficult to grow, such as Legionella species, may reduce difficulties encountered with culture and immunofluorescent staining. We evaluated a commercial PCR and hybridization kit, designed for environmental samples, for the detection of Legionella in respiratory specimens. Sixteen Legionella species cultures tested positive with the Perkin Elmer Legionella EnviroAmp Amplification and Detection kits (Perkin Elmer, Foster City, Calif)...
September 1998: American Journal of Clinical Pathology
C P Saint
Modifications to the EnviroAmp Legionella detection system are described which permit the rapid analysis of bacterial colonies taken from Legionella selective media. Capillary PCR permitted twice the number of samples to be analysed with a single kit. When PCR was positive for Leg. pneumophila, this result was confirmed by seroagglutination. The reverse dot blot hybridization assay was only used where PCR indicated a Legionella sp. other than Leg. pneumophila, permitting further savings on detection system components...
May 1998: Letters in Applied Microbiology
V Catalan, F Garcia, C Moreno, M J Vila, D Apraiz
The study of Legionella in treated wastewater acquires special importance when this water is used in irrigation by spray, as Legionella is transmitted via the inhalation of aerosols and may consequently represent a health risk. In this study, we applied polymerase chain reaction (PCR) amplification as an alternative method to plate culture for detecting L. pneumophila in twelve heavily biocontaminated samples from a wastewater treatment plant. Moreover, we studied the efficiency of rapid gel filtration methods and filtration through chelating ion exchange resin in the elimination of PCR inhibitors from wastewater samples...
January 1997: Research in Microbiology
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