Articoli di riviste: "Legionella detection"

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Cloud, J. L., K. C. Carroll, P. Pixton, M. Erali e D. R. Hillyard. "Detection of Legionella Species in Respiratory Specimens Using PCR with Sequencing Confirmation". Journal of Clinical Microbiology 38, n. 5 (2000): 1709–12. http://dx.doi.org/10.1128/jcm.38.5.1709-1712.2000.

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Legionella spp. are a common cause of community-acquired respiratory tract infections and an occasional cause of nosocomial pneumonia. A PCR method for the detection of legionellae in respiratory samples was evaluated and was compared to culture. The procedure can be performed in 6 to 8 h with a commercially available DNA extraction kit (Qiagen, Valencia, Calif.) and by PCR with gel detection. PCR is performed with primers previously determined to amplify a 386-bp product within the 16S rRNA gene of Legionella pneumophila. We can specifically detect the clinically significant Legionella species including L. pneumophila, L. micdadei, L. longbeachae, L. bozemanii, L. feeleii, and L. dumoffii. The assay detects 10 fg (approximately two organisms) of legionella DNA in each PCR. Of 212 clinical specimens examined by culture, 100% of the culture-positive samples (31 of 31) were positive by this assay. By gel detection of amplification products, 12 of 181 culture-negative samples were positive forLegionella species by PCR, resulting in 93% specificity. Four of the 12 samples with discrepant results (culture negative, PCR positive) were confirmed to be positive for Legionellaspecies by sequencing of the amplicons. The legionella-specific PCR assay that is described demonstrates high sensitivity and high specificity for routine detection of legionellae in respiratory samples.

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Toplitsch, Daniela, Sabine Platzer, Romana Zehner, Stephanie Maitz, Franz Mascher e Clemens Kittinger. "Comparison of Updated Methods for Legionella Detection in Environmental Water Samples". International Journal of Environmental Research and Public Health 18, n. 10 (19 maggio 2021): 5436. http://dx.doi.org/10.3390/ijerph18105436.

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The difficulty of cultivation of Legionella spp. from water samples remains a strenuous task even for experienced laboratories. The long incubation periods for Legionellae make isolation difficult. In addition, the water samples themselves are often contaminated with accompanying microbial flora, and therefore require complex cultivation methods from diagnostic laboratories. In addition to the recent update of the standard culture method ISO 11731:2017, new strategies such as quantitative PCR (qPCR) are often discussed as alternatives or additions to conventional Legionella culture approaches. In this study, we compared ISO 11731:2017 with qPCR assays targeting Legionella spp., Legionella pneumophila, and Legionella pneumophila serogroup 1. In samples with a high burden of accompanying microbial flora, qPCR shows an excellent negative predictive value for Legionella pneumophila, thus making qPCR an excellent tool for pre-selection of negative samples prior to work-intensive culture methods. This and its low limit of detection make qPCR a diagnostic asset in Legionellosis outbreak investigations, where quick-risk assessments are essential, and are a useful method for monitoring risk sites.

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Fricker, E. J., e C. R. Fricker. "Detection of legionella spp. using a commercially available polymerase chain reaction test". Water Science and Technology 31, n. 5-6 (1 marzo 1995): 407–8. http://dx.doi.org/10.2166/wst.1995.0649.

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A commercial test kit (the EnviroAmp Legionella Kit) for the detection of legionellas using the polymerase chain reaction is compared with the standard culture methods for water samples. The EnviroAmp kit proved to be rapid and did not require great experience of molecular biological techniques. However as number of samples which tested negative with the standard culture, were positive with the kit; further research is needed to establish whether this due to the detection of dead or viable but non-culturable legionellas.

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Wellinghausen, Nele, Cathrin Frost e Reinhard Marre. "Detection of Legionellae in Hospital Water Samples by Quantitative Real-Time LightCycler PCR". Applied and Environmental Microbiology 67, n. 9 (1 settembre 2001): 3985–93. http://dx.doi.org/10.1128/aem.67.9.3985-3993.2001.

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ABSTRACT Contamination of hospital water systems with legionellae is a well-known cause of nosocomial legionellosis. We describe a new real-time LightCycler PCR assay for quantitative determination of legionellae in potable water samples. Primers that amplify both a 386-bp fragment of the 16S rRNA gene from Legionellaspp. and a specifically cloned fragment of the phage lambda, added to each sample as an internal inhibitor control, were used. The amplified products were detected by use of a dual-color hybridization probe assay design and quantified with external standards composed ofLegionella pneumophila genomic DNA. The PCR assay had a sensitivity of 1 fg of Legionella DNA (i.e., less than one Legionella organism) per assay and detected 44Legionella species and serogroups. Seventy-seven water samples from three hospitals were investigated by PCR and culture. The rates of detection of legionellae were 98.7% (76 of 77) by the PCR assay and 70.1% (54 of 77) by culture; PCR inhibitors were detected in one sample. The amounts of legionellae calculated from the PCR results were associated with the CFU detected by culture (r= 0.57; P < 0.001), but PCR results were mostly higher than the culture results. Since L. pneumophila is the main cause of legionellosis, we further developed a quantitativeL. pneumophila-specific PCR assay targeting the macrophage infectivity potentiator (mip) gene, which codes for an immunophilin of the FK506 binding protein family. All but one of the 16S rRNA gene PCR-positive water samples were also positive in the mip gene PCR, and the results of the two PCR assays were correlated. In conclusion, the newly developedLegionella genus-specific and L. pneumophila species-specific PCR assays proved to be valuable tools for investigation of Legionella contamination in potable water systems.

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Chambers, Stephen T., Sandy Slow, Amy Scott-Thomas e David R. Murdoch. "Legionellosis Caused by Non-Legionella pneumophila Species, with a Focus on Legionella longbeachae". Microorganisms 9, n. 2 (31 gennaio 2021): 291. http://dx.doi.org/10.3390/microorganisms9020291.

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Although known as causes of community-acquired pneumonia and Pontiac fever, the global burden of infection caused by Legionella species other than Legionella pneumophila is under-recognised. Non-L. pneumophila legionellae have a worldwide distribution, although common testing strategies for legionellosis favour detection of L. pneumophila over other Legionella species, leading to an inherent diagnostic bias and under-detection of cases. When systematically tested for in Australia and New Zealand, L. longbeachae was shown to be a leading cause of community-acquired pneumonia. Exposure to potting soils and compost is a particular risk for infection from L. longbeachae, and L. longbeachae may be better adapted to soil and composting plant material than other Legionella species. It is possible that the high rate of L. longbeachae reported in Australia and New Zealand is related to the composition of commercial potting soils which, unlike European products, contain pine bark and sawdust. Genetic studies have demonstrated that the Legionella genomes are highly plastic, with areas of the chromosome showing high levels of recombination as well as horizontal gene transfer both within and between species via plasmids. This, combined with various secretion systems and extensive effector repertoires that enable the bacterium to hijack host cell functions and resources, is instrumental in shaping its pathogenesis, survival and growth. Prevention of legionellosis is hampered by surveillance systems that are compromised by ascertainment bias, which limits commitment to an effective public health response. Current prevention strategies in Australia and New Zealand are directed at individual gardeners who use potting soils and compost. This consists of advice to avoid aerosols generated by the use of potting soils and use masks and gloves, but there is little evidence that this is effective. There is a need to better understand the epidemiology of L. longbeachae and other Legionella species in order to develop effective treatment and preventative strategies globally.

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Aoki, S., Y. Hirakata, Y. Miyazaki, K. Izumikawa, K. Yanagihara, K. Tomono, Y. Yamada, T. Tashiro, S. Kohno e S. Kamihira. "Detection of Legionella DNA by PCR of whole-blood samples in a mouse model". Journal of Medical Microbiology 52, n. 4 (1 aprile 2003): 325–29. http://dx.doi.org/10.1099/jmm.0.04999-0.

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A detection system for Legionella DNA in blood samples based on the PCR was developed and evaluated in A/J mice with experimentally induced Legionella pneumonia. Primers were designed to amplify a 106 bp DNA fragment of the 16S rRNA gene specific to Legionella species. The PCR system could detect clinically relevant Legionella species including Legionella pneumophila, Legionella micdadei, Legionella bozemanae, Legionella dumoffii, Legionella longbeachae, Legionella gormanii and Legionella jordanis. The sensitivity of the PCR system was 20 fg extracted DNA. In the mouse model, the blood PCR was compared with results obtained by PCR on bronchoalveolar lavage fluid (BALF) samples, cultures of blood and BALF and detection of Legionella urinary antigen. Blood PCR was positive until 8 days after infection, while BALF PCR became negative on day 4. These results indicate that PCR using blood samples may be a useful, convenient and non-invasive method for the diagnosis of Legionella pneumonia.

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Albalat, Guillermo Rodríguez, Begoña Bedrina Broch e Marisa Jiménez Bono. "Method Modification of the Legipid®Legionella Fast Detection Test Kit". Journal of AOAC INTERNATIONAL 97, n. 5 (1 settembre 2014): 1403–9. http://dx.doi.org/10.5740/jaoacint.14-029.

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Abstract Legipid®Legionella Fast Detection is a test based on combined magnetic immunocapture and enzyme-immunoassay (CEIA) for the detection of Legionella in water. The test is based on the use of anti-Legionella antibodies immobilized on magnetic microspheres. Target microorganism is preconcentrated by filtration. Immunomagnetic analysis is applied on these preconcentrated water samples in a final test portion of 9 mL. The test kit was certified by the AOAC Research Institute as Performance Tested MethodSM (PTM) No. 111101 in a PTM validation which certifies the performance claims of the test method in comparison to the ISO reference method 11731-1998 and the revision 11731-2004 “Water Quality: Detection and Enumeration of Legionella pneumophila” in potable water, industrial water, and waste water. The modification of this test kit has been approved. The modification includes increasing the target analyte from L. pneumophila to Legionella species and adding an optical reader to the test method. In this study, 71 strains of Legionella spp. other than L. pneumophila were tested to determine its reactivity with the kit based on CEIA. All the strains of Legionella spp. tested by the CEIA test were confirmed positive by reference standard method ISO 11731. This test (PTM 111101) has been modified to include a final optical reading. A methods comparison study was conducted to demonstrate the equivalence of this modification to the reference culture method. Two water matrixes were analyzed. Results show no statistically detectable difference between the test method and the reference culture method for the enumeration of Legionella spp. The relative level of detection was 93 CFU/volume examined (LOD50). For optical reading, the LOD was 40 CFU/volume examined and the LOQ was 60 CFU/volume examined. Results showed that the test Legipid Legionella Fast Detection is equivalent to the reference culture method for the enumeration of Legionella spp.

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Oshiro, Robin K., Teresa Picone e Betty H. Olson. "Modification of reagents in the EnviroAmp™ kit to increase recovery of Legionella organisms in water". Canadian Journal of Microbiology 40, n. 6 (1 giugno 1994): 495–99. http://dx.doi.org/10.1139/m94-080.

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Organisms of the bacterial genus Legionella, commonly found in aqueous reservoirs, have been associated with Legionnaires' disease (legionella pneumonia, caused by Legionella pneumophila) and Pontiac fever (nonpneumonic legionellosis). EnviroAmp™ Legionella sample preparation, polymerase chain reaction amplification, and detection kits (Perkin-Elmer Corp.) were developed for rapid detection of DNA from organisms of the genus Legionella and the species L. pneumophila from environmental water samples. The kits are based on molecular techniques incorporating polymerase chain reaction amplification and detection by reverse dot blot hybridization to particular genus and species probes. The manufacturer states that the EnviroAmp™ Legionella sample preparation, polymerase chain reaction amplification, and detection kits can detect approximately 100 Legionella organisms/mL (10 000 organisms/100 mL) in the original water sample. The sensitivity of the kits was increased to 0.1 colony-forming units/mL (10 colony-forming units/100 mL), at least for cultured organisms, by modifying the EnviroAmp™ Legionella sample preparation kit protocol. Data obtained in this study indicated that sample volume could be increased from 100 to 1000 mL (in the absence of interfering substances such as humic acid) and DNA extraction volume could be decreased from 2 to 0.5 mL to increase the ability of the kit to detect lower numbers of Legionella spp. or L. pneumophila per volume.Key words: Legionella, environment, water, EnviroAmp™ kit.

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Chang, Bin, Kanji Sugiyama, Toshitsugu Taguri, Junko Amemura-Maekawa, Fumiaki Kura e Haruo Watanabe. "Specific Detection of Viable Legionella Cells by Combined Use of Photoactivated Ethidium Monoazide and PCR/Real-Time PCR". Applied and Environmental Microbiology 75, n. 1 (31 ottobre 2008): 147–53. http://dx.doi.org/10.1128/aem.00604-08.

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ABSTRACT Legionella organisms are prevalent in manmade water systems and cause legionellosis in humans. A rapid detection method for viable Legionella cells combining ethidium monoazide (EMA) and PCR/real-time PCR was assessed. EMA could specifically intercalate and cleave the genomic DNA of heat- and chlorine-treated dead Legionella cells. The EMA-PCR assay clearly showed an amplified fragment specific for Legionella DNA from viable cells, but it could not do so for DNA from dead cells. The number of EMA-treated dead Legionella cells estimated by real-time PCR exhibited a 104- to 105-fold decrease compared to the number of dead Legionella cells without EMA treatment. Conversely, no significant difference in the numbers of EMA-treated and untreated viable Legionella cells was detected by the real-time PCR assay. The combined assay was also confirmed to be useful for specific detection of culturable Legionella cells from water samples obtained from spas. Therefore, the combined use of EMA and PCR/real-time PCR detects viable Legionella cells rapidly and specifically and may be useful in environmental surveillance for Legionella.

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Roll, Bruce M., e Roger S. Fujioka. "Detection of legionella bacteria in sewage by polymerase chain reaction and standard culture method". Water Science and Technology 31, n. 5-6 (1 marzo 1995): 409–16. http://dx.doi.org/10.2166/wst.1995.0650.

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Legionella bacteria are ubiquitous in environmental waters. Only a few species of Legionella , especially, L. pneumophila are pathogenic to humans and cause a sometimes fatal Legionnaires disease as well as a less fatal disease called Pontiac fever. The presence of Legionella in sewage and aerosolized sewage is the subject of this investigation because reuse of sewage may involve the exposure of people to aerosolization, the mode of transmission of Legionella bacteria. The objective of this study was to determine the prevalence of Legionella species and L. pneumophila in wastewater and their fate after various stages of treatment. The polymerase chain reaction (PCR) and standard culture method were utilized to detect Legionella species and L. pneumophila. PCR results indicated that Legionella species were present at levels > 103 cells / ml during all phases of sewage treatment including chlorinated effluents. Culture results indicated levels at least one log lower than seen with PCR. Legionella species were also recovered from air samples collected from secondary aeration basins at levels < 103 cells/ml. PCR was shown to be the most rapid and sensitive method for detecting Legionella in sewage.

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Martín Pérez, Leonardo, Francesc Codony, Karina Ríos, Bárbara Adrados, Mariana Fittipaldi, Gregori De Dios, Gustavo Peñuela e Jordi Morató. "Prevalence study of Simkania negevensis in cooling towers in Spain". Journal of Water and Health 9, n. 2 (23 dicembre 2010): 312–16. http://dx.doi.org/10.2166/wh.2010.216.

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Simkania negevensis is an obligate intracellular bacterium grouped into the order Chlamydiales. This new amoeba-resistant intracellular bacterium might represent a novel etiologic agent of bronchiolitis and community-acquired pneumonia and occurs in aquatic habitats such as drinking water and reclaimed wastewater. Another amoeba-related bacterium, Legionella pneumophila, is an etiologic agent of pneumonia transmitted by environmental aerosols or contaminated water/air cooling systems. These transmission pathways are important in the natural history of Legionellae infections and possibly other intracellular microorganisms such as Parachlamydiaceae; thus, understanding the feasibility of Simkania infection by these routes is relevant. In the present work, we investigated the prevalence of this newly identified pathogenic bacterium in cooling towers by quantitative PCR (qPCR) and its possible relationship with Legionella pneumophila co-infection. Our results show Simkania detection in 2 of 70 cooling towers analyzed. To our knowledge, this report is the first describing Simkania negevensis detection in this category of environmental water samples.

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Wong, Alicia Y. W., Alexander T. A. Johnsson, Aina Iversen, Simon Athlin e Volkan Özenci. "Evaluation of Four Lateral Flow Assays for the Detection of Legionella Urinary Antigen". Microorganisms 9, n. 3 (26 febbraio 2021): 493. http://dx.doi.org/10.3390/microorganisms9030493.

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Urinary antigen tests (UATs) are often used to diagnose Legionnaires’ disease as they are rapid and easy to perform on readily obtainable urine samples without the need for specialized skills compared to conventional methods. Recently developed automated readers for UATs may provide objective results interpretation, especially in cases of weak result bands. Using 53 defined patient urine samples, we evaluated the performance of the BinaxNOW Legionella Antigen Card (Abbott), ImmuView S. pneumoniae and Legionella (SSI Diagnostica), STANDARD F Legionella Ag FIA (SD Biosensor), and Sofia Legionella FIA (Quidel) simultaneously with their respective automated readers. Automatic and visual interpretation of result bands were also compared for the immunochromatography-based BinaxNOW and ImmuView UATs. Overall sensitivity and specificity of Legionella UATs were 53.9–61.5% and 90.0–94.9%, respectively. All four UATs successfully detected all samples from L. pneumophila serogroup 1-positive patients, but most failed to detect samples for Legionella spp., or other serogroups. Automatic results interpretation of results was found to be mostly concordant with visual results reading. In conclusion, the performance of the four UATs were similar to each other in the detection of Legionella urinary antigen with no major difference between automated or visual results reading.

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Tateda, Kazuhiro. "Legionella Pneumonia". Journal of Disaster Research 6, n. 4 (1 agosto 2011): 435–42. http://dx.doi.org/10.20965/jdr.2011.p0435.

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Legionella pneumonia, an infectious disease with high fatality, produces symptoms after inhalation of an aerosol contaminated by Legionella species. Legionella bacteria are Gram-negative, glucose-nonfermentative, and proliferative in human macrophages and monocytes. Over 40 types of Legionella species are pathogenic in human beings. Of these,L. pneumophilahas the strongest pathogenicity. Hot springs and bathtubs with circulating systems are considered sources of infection and sometimes cause cases of hospital-acquired infection. The urinary antigen detection test is a new diagnostic technique, and proved to be effective for rapid and correct diagnosis ofL. pneumophilaserogroup-1 infection.

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Declerck, P., L. Verelst, L. Duvivier, A. Van Damme e F. Ollevier. "A detection method for Legionella spp in (cooling) water: fluorescent in situ hybridisation (FISH) on whole bacteria". Water Science and Technology 47, n. 3 (1 febbraio 2003): 143–46. http://dx.doi.org/10.2166/wst.2003.0184.

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Although traditional culture methods are appropriate for detection of Legionella species, such culture takes several days. Rapid detection (<24 h) of individual Legionella is possible using fluorescent in situ hybridisation (FISH) on whole bacteria. Water samples were filtered and the concentrated bacteria were immediately detected (without culture) with a fluorescence microscope following appropriate labelling. The detection level was very high and quantification was possible. For the detection of all Legionella spp. the probe LEG705 was used, complementary to a 16S rRNA sequence conserved in all Legionella spp. For specific detection of L. pneumophila the probe LEGPNE1 was used. This probe is designed against a variable domain of the 16S rRNA sequence from L. pneumophila. CY3 and FLUOS labels were tested and CY3 showed clearly detectable bacteria with minimum background staining. This FISH technique is very sensitive, fast, reliable and individual bacteria are easily detected.

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Párraga-Niño, Noemí, Sara Quero, Naroa Uria, Oscar Castillo-Fernandez, Josune Jimenez-Ezenarro, Francesc-Xavier Muñoz, Miquel Sabrià e Marian Garcia-Nuñez. "Antibody test for Legionella pneumophila detection". Diagnostic Microbiology and Infectious Disease 90, n. 2 (febbraio 2018): 85–89. http://dx.doi.org/10.1016/j.diagmicrobio.2017.11.005.

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Delgado-Viscogliosi, Pilar, Tristan Simonart, Virginie Parent, Grégory Marchand, Marie Dobbelaere, Eric Pierlot, Véronique Pierzo et al. "Rapid Method for Enumeration of Viable Legionella pneumophila and Other Legionella spp. in Water". Applied and Environmental Microbiology 71, n. 7 (luglio 2005): 4086–96. http://dx.doi.org/10.1128/aem.71.7.4086-4096.2005.

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ABSTRACT A sensitive and specific method has been developed to enumerate viable L. pneumophila and other Legionella spp. in water by epifluorescence microscopy in a short period of time (a few hours). This method allows the quantification of L. pneumophila or other Legionella spp. as well as the discrimination between viable and nonviable Legionella. It simultaneously combines the specific detection of Legionella cells using antibodies and a bacterial viability marker (ChemChrome V6), the enumeration being achieved by epifluorescence microscopy. The performance of this immunological double-staining (IDS) method was investigated in 38 natural filterable water samples from different aquatic sources, and the viable Legionella counts were compared with those obtained by the standard culture method. The recovery rate of the IDS method is similar to, or higher than, that of the conventional culture method. Under our experimental conditions, the limit of detection of the IDS method was <176 Legionella cells per liter. The examination of several samples in duplicates for the presence of L. pneumophila and other Legionella spp. indicated that the IDS method exhibits an excellent intralaboratory reproducibility, better than that of the standard culture method. This immunological approach allows rapid measurements in emergency situations, such as monitoring the efficacy of disinfection shock treatments. Although its field of application is as yet limited to filterable waters, the double-staining method may be an interesting alternative (not equivalent) to the conventional standard culture methods for enumerating viable Legionella when rapid detection is required.

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Asghari, Farzaneh Baghal, Mahnaz Nikaeen, Maryam Hatamzadeh e Akbar Hassanzadeh. "Surveillance of Legionella species in hospital water systems: the significance of detection method for environmental surveillance data". Journal of Water and Health 11, n. 4 (12 settembre 2013): 713–19. http://dx.doi.org/10.2166/wh.2013.064.

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Monitoring of hospital water systems to prevent and control nosocomial legionellosis is important from a public health perspective. This study was conducted to survey the prevalence of Legionella contamination of hospital waters. A total of 44 water samples from the hot-water system of 11 hospitals were tested for Legionella by a culture method and a nested polymerase chain reaction (PCR) assay with Legionella-specific primers to identify the more sensitive method. Some physicochemical parameters and heterotrophic plate counts of water samples for possible association with Legionella contamination were also determined. The contamination rate of hospitals in our study varied between 64% (eight of 11)–100% based on culture method and nested PCR, respectively. Of the 44 water samples examined, 23% were positive for Legionella spp. by the culture method, while the nested PCR assay using the primers LEG448-JRP revealed 66% of the water samples being positive. Given the importance of monitoring hospital water systems for the presence of Legionella spp., the present PCR assay proved highly applicable for practical and sensitive surveillance of Legionella in such water systems. In addition, rapid monitoring of Legionella contamination could eliminate the potential exposure of high-risk patients through effective control measures.

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Lund, Vidar, Wenche Fonahn, Jens Erik Pettersen, Dominique A. Caugant, Eirik Ask e Åse Nysaeter. "Detection of Legionella by cultivation and quantitative real-time polymerase chain reaction in biological waste water treatment plants in Norway". Journal of Water and Health 12, n. 3 (31 gennaio 2014): 543–54. http://dx.doi.org/10.2166/wh.2014.063.

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Cases of Legionnaires' disease associated with biological treatment plants (BTPs) have been reported in six countries between 1997 and 2010. However, knowledge about the occurrence of Legionella in BTPs is scarce. Hence, we undertook a qualitative and quantitative screening for Legionella in BTPs treating waste water from municipalities and industries in Norway, to assess the transmission potential of Legionella from these installations. Thirty-three plants from different industries were sampled four times within 1 year. By cultivation, 21 (16%) of 130 analyses were positive for Legionella species and 12 (9%) of 130 analyses were positive for Legionella pneumophila. By quantitative real-time polymerase chain reaction (PCR), 433 (99%) of 437 analyses were positive for Legionella species and 218 (46%) of 470 analyses were positive for L. pneumophila. This survey indicates that PCR could be the preferable method for detection of Legionella in samples from BTPs. Sequence types of L. pneumophila associated with outbreaks in Norway were not identified from the BTPs. We showed that a waste water treatment plant with an aeration basin can produce high concentrations of Legionella. Therefore, these plants should be considered as a possible source of community-acquired Legionella infections.

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Kümpers, Philipp, Andreas Tiede, Philip Kirschner, Jutta Girke, Arnold Ganser e Dietrich Peest. "Legionnaires' disease in immunocompromised patients: a case report of Legionella longbeachae pneumonia and review of the literature". Journal of Medical Microbiology 57, n. 3 (1 marzo 2008): 384–87. http://dx.doi.org/10.1099/jmm.0.47556-0.

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In addition to Legionella pneumophila, about 20 Legionella species have been documented as human pathogens. The majority of infections by non-pneumophila Legionella species occur in immunocompromised and splenectomized patients. Here, we report a case of ‘classical’ lobar pneumonia caused by Legionella longbeachae in a splenectomized patient receiving corticosteroids for chronic immune thrombocytopenia. Tests for Legionella antigen were negative. L. longbeachae was immediately detected in bronchoalveolar fluid by PCR and subsequently confirmed by culture on legionella-selective media. The features of Legionnaires' disease in immunocompromised patients with special emphasis on significance and detection of non-pneumophila species are reviewed.

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Muchesa, P., M. Lelfels, L. Jurzik, T. G. Barnard e C. Bartle. "Detection of amoeba-associated Legionella pneumophila in hospital water networks of Johannesburg". Southern African Journal of Infectious Diseases 33, n. 3 (30 settembre 2018): 72–75. http://dx.doi.org/10.4102/sajid.v33i3.8.

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The prevalence of free-living amoeba and associated Legionella spp. in hospital water systems may pose a risk of Legionnaires’ disease to immuno-compromised patients. This study investigated the occurrence of amoeba-associated Legionella pneumophila in three South African hospital water systems. A total of 98 water and/or biofilm samples were collected from the sterilisation unit, theatres, neonatal ward and intensive care units. Amoebae were isolated from 71 (72.4%) samples. Isolated amoebae were analysed using qPCR and culture methods to test for the presence of Legionella. L. pneumophila did not grow on selective media in any of the samples. A total of 7 out of the 71 (9.9%) amoeba-positive samples showed a positive reaction for L. pneumophila using qPCR. Although relatively few samples were positive for Legionella in this preliminary study, the association with amoeba still presents a potential public health risk to immuno-compromised patients when exposed to contaminated water.

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Ditommaso, Savina, Monica Giacomuzzi, Gabriele Memoli, Jacopo Garlasco e Carla M. Zotti. "Sensitivity and Selectivity of Two Commercially Available Media for Legionella spp. Recovery from Environmental Water Samples". Pathogens 9, n. 7 (29 giugno 2020): 523. http://dx.doi.org/10.3390/pathogens9070523.

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The quality control of culture media used for Legionella spp. isolation and enumeration is paramount to achieve a satisfactory degree of comparability among water testing results from different laboratories. Here, we report on a comparative assessment of the sensitivity and selectivity of MWY and BCYEα media supplied by two different manufacturers (i.e., Xebios Diagnostics GmbH and Oxoid Ltd) for the detection of Legionella spp. from environmental water samples. Even though our analysis showed an excellent agreement between the recovery rates of the four media tested (90.5%), the quantitative recovery of Legionella spp. colonies using Xebios media was significantly greater than that achieved by Oxoid media (P = 0.0054). Furthermore, the sensitivity of detection was significantly higher when samples were plated on MWY Xebios agar (P = 0.0442), while the selectivity of MWY appeared to be the same regardless of the manufacturer. Furthermore, MWYXebios agar favored the growth of much larger colonies compared to those observed on MWYOxoid agar. Finally, MWYXebios medium enhanced the recovery of non-pneumophila Legionella species. Collectively, our findings demonstrate that quality control is crucial to ensure high selectivity and sensitivity of the culture media used for the detection and enumeration of Legionella spp. from environmental water resources. As water remediation measures strictly depend on Legionella spp. recovery, culture protocol standardization, as well as quality control of the culture media, is essential to achieve intra- and interlaboratory reproducibility and accuracy.

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JIBIKI, Kazuko, Saeko NAKAMURA, Satoshi OOI, Teppei KUMADA, Reiko DEMURA, Emi ODAGIRI, Hiroshi DEMURA, Kihachiro SHIMIZU e Toshiharu YAMAMORI. "Detection of Urinary Legionella Antigen by Radioimmunoassay". Journal of the Japanese Association for Infectious Diseases 59, n. 1 (1985): 1–8. http://dx.doi.org/10.11150/kansenshogakuzasshi1970.59.1.

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KUMADA, Teppei, Naoki TOMORI, Keizoh KAZONO, Reiko DEMURA, Emi ODAGIRI, Satoshi OOI, Hiroshi DEMURA et al. "Detection of Urinary Legionella Antigen by Radioimmunoassay". Journal of the Japanese Association for Infectious Diseases 59, n. 1 (1985): 9–13. http://dx.doi.org/10.11150/kansenshogakuzasshi1970.59.9.

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24

Mietzner, Sue M., e Janet E. Stout. "Laboratory detection of Legionella in environmental samples". Clinical Microbiology Newsletter 24, n. 11 (giugno 2002): 81–85. http://dx.doi.org/10.1016/s0196-4399(02)80017-4.

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25

Schwaminger, Sebastian, Marina E. Rottmueller, Ramona Fischl, Behnam Kalali e Sonja Berensmeier. "Detection of targeted bacteria species on filtration membranes". Analyst 146, n. 11 (2021): 3549–56. http://dx.doi.org/10.1039/d1an00117e.

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26

Kessler, H. H., F. F. Reinthaler, A. Pschaid, K. Pierer, B. Kleinhappl, E. Eber e E. Marth. "Rapid detection of Legionella species in bronchoalveolar lavage fluids with the EnviroAmp Legionella PCR amplification and detection kit." Journal of Clinical Microbiology 31, n. 12 (1993): 3325–28. http://dx.doi.org/10.1128/jcm.31.12.3325-3328.1993.

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27

He, Ying, Tsung C. Chang, Haijing Li, Gongyi Shi e Yi-Wei Tang. "Matrix-assisted laser desorption ionization time-of-flight mass spectrometry and database for identification of Legionella species 1This study was presented in part at the 110th American Society for Microbiology Annual Meeting, 23–27 May 2010, San Diego, California." Canadian Journal of Microbiology 57, n. 7 (luglio 2011): 533–38. http://dx.doi.org/10.1139/w11-039.

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Abstract (sommario):

More than 20 species of Legionella have been identified in relation to human infections. Rapid detection and identification of Legionella isolates is clinically useful to differentiate between infection and contamination and to determine treatment regimens. We explored the use of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) Biotyper system (Bruker Daltonik GmbH, Bremen, Germany) for the identification of Legionella species. The MALDI MS spectra were generated and compared with the Biotyper database, which includes 25 Legionella strains covering 22 species and four Legionella pneumophila serogroups. A total of 83 blind-coded Legionella strains, consisting of 54 reference and 29 clinical strains, were analyzed in the study. Overall, the Biotyper system correctly identified 51 (61.4%) of all strains and isolates to the species level. For species included in the Biotyper database, the method identified 51 (86.4%) strains out of 59 Legionella strains to the correct species level, including 24 (100%) L. pneumophila and 27 (77.1%) non-L. pneumophila strains. The remaining 24 Legionella strains, belonging to species not covered by the Biotyper database, were either identified to the Legionella genus level or had no reliable identification. The Biotyper system produces constant and reproducible MALDI MS spectra for Legionella strains and can be used for rapid and accurate Legionella identification. More Legionella strains, especially the non-L. pneumophila strains, need to be included in the current Biotyper database to cover varieties of Legionella species and to increase identification accuracy.

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Scaturro, Maria, Elisa Poznanski, Mariarosaria Mupo, Paola Blasior, Margit Seeber, Anna-Maria Prast, Elisa Romanin et al. "Evaluation of GVPC and BCYE Media for Legionella Detection and Enumeration in Water Samples by ISO 11731: Does Plating on BCYE Medium Really Improve Yield?" Pathogens 9, n. 9 (16 settembre 2020): 757. http://dx.doi.org/10.3390/pathogens9090757.

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Abstract (sommario):

Legionella spp are the causative agents of Legionnaires’ diseases, which is a pneumonia of important public health concern. Ubiquitous freshwater and soil inhabitants can reach man-made water systems and cause illness. Legionella enumeration and quantification in water systems is crucial for risk assessment and culture examination is the gold standard method. In this study, Legionella recovery from potable water samples, at presumably a low concentration of interfering microorganisms, was compared by plating on buffered charcoal yeast extract (BCYE) and glycine, vancomycin, polymyxin B, cycloheximide (GVPC) Legionella agar media, according to the International Standard Organization (ISO) 11731: 2017. Overall, 556 potable water samples were analyzed and 151 (27.1%) were positive for Legionella. Legionella grew on both BCYE and GVPC agar plates in 85/151 (56.3%) water samples, in 65/151 (43%) on only GVPC agar plates, and in 1/151 (0.7%) on only BCYE agar plates. In addition, GVPC medium identified Legionella species other than pneumophila in six more samples as compared with the culture on BCYE. Although the medians of colony forming units per liter (CFU/L) detected on the BCYE and GVPC agar plates were 2500 and 1350, respectively (p-value < 0.0001), the difference did not exceed one logarithm, and therefore is not relevant for Legionella risk assessment. These results make questionable the need to utilize BCYE agar plates to analyze potable water samples.

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Su, Han, Shaopei Li, Mauricio Terebiznik, Cyril Guyard e Kagan Kerman. "Biosensors for the Detection of Interaction between Legionella pneumophila Collagen-Like Protein and Glycosaminoglycans". Sensors 18, n. 8 (14 agosto 2018): 2668. http://dx.doi.org/10.3390/s18082668.

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The adhesin Legionella collagen-like (Lcl) protein can bind to extracellular matrix components and mediate the binding of Legionella pneumophila to host cells. In this study, electrochemical impedance spectroscopy (EIS) and surface plasmon resonance (SPR)-based biosensors were employed to characterize these interactions between glycosaminoglycans (GAGs) and the adhesin Lcl protein. Fucoidan displayed a high affinity (KD 18 nM) for Lcl protein. Chondroitin sulfate A and dermatan sulfate differ in the position of a carboxyl group replacing D-glucuronate with D-iduronate. Our results indicated that the presence of D-iduronate in dermatan sulfate strongly hindered its interaction with Lcl. These biophysical studies provided valuable information in our understanding of adhesin-ligand interactions related to Legionella pneumophila infections.

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De Giglio, Osvalda, Giusy Diella, Paolo Trerotoli, Michela Consonni, Roberta Palermo, Marina Tesauro, Pasqualina Laganà, Gabriella Serio e Maria Teresa Montagna. "Legionella Detection in Water Networks as per ISO 11731:2017: Can Different Filter Pore Sizes and Direct Placement on Culture Media Influence Laboratory Results?" International Journal of Environmental Research and Public Health 17, n. 6 (20 marzo 2020): 2077. http://dx.doi.org/10.3390/ijerph17062077.

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Determination of Legionella concentrations in water networks is useful for predicting legionellosis risks. The standard culture technique using concentration with membranes filters is the most commonly used method for environmental surveillance of Legionella. The aim of this study was to verify whether filtration with different filter pore sizes (0.2 and 0.45 µm) according to (ISO) 11731:2017, followed by directly placing them on culture media, can influence Legionella detection. Three laboratories participated in an experimental study that tested a known suspension of Legionella pneumophila (Lpn) serogroup 1 (ATCC 33152) (approximate final cell density of 15 CFU/mL). E. coli (ATCC 11775) and Pseudomonas aeruginosa (ATCC 25668) were included as control tests. The average (95% CI) percentage of recovery of Lpn was 65% using 0.45-µm filters and 15% using 0.2-µm filters (p < 0.0001). For control tests, the average (95% CI) percentage of recovery was higher with 0.45 vs. 0.2 µm filters: 97% vs. 64% for Escherichia coli (p < 0.00001) and 105% vs. 97% (p = 0.0244) for P. aeruginosa. Our results showed that the 0.45-µm filters provided the greatest detection of Legionella. Because the current national guidelines leave the choice of membrane porosity to the operator, experimental studies are important for directing operators towards a conscious choice to standardize Legionella environmental surveillance methods.

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Yarom, Rachel, Rivka Sheinman e Robert Armon. "Legionella pneumophila serogroup 3 prevalence in drinking water survey in Israel (2003–2007)". Water Supply 10, n. 5 (1 dicembre 2010): 746–52. http://dx.doi.org/10.2166/ws.2010.489.

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Abstract (sommario):

A 5-year survey (2003 to 2007) was carried out on various drinking water sources in Israel for detection of Legionella spp. The most frequently isolated species was L. pneumophila serogroup 3 (prevalence 60.44±5.6) followed by L. pneumophila serogroup 1 (26.98±5.2) and other serogroups and unidentified species (12.6±9.8). L. pneumophila serogroup 1 is the worldwide prevalent serogroup in water sources while serogroup 3 was only occasionally reported. Previous reports on L. pneumophila serogroup 1 as the main causative of respiratory disease in Israel revealed a drop from 52 to 15% in incidence during a 5-year survey, with a rise in the incidence of seropositivity to “other Legionellae”, mainly sg. 3. As antigenuria is the main clinical tool to detect Legionellosis among infected patients, the shift from serogroup 1 to serogroup 3 in water sources will go undetected for obvious reasons. The process of serogroups prevalence shift in water sources is an interesting issue that has to be investigated further in order to advance our understanding of Legionella epidemiology.

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Thomas, Julian A., Florian Schnell, Yasmin Kaveh-Baghbaderani, Sonja Berensmeier e Sebastian P. Schwaminger. "Immunomagnetic Separation of Microorganisms with Iron Oxide Nanoparticles". Chemosensors 8, n. 1 (27 febbraio 2020): 17. http://dx.doi.org/10.3390/chemosensors8010017.

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The early detection of Legionella in water reservoirs, and the prevention of their often fatal diseases, requires the development of rapid and reliable detection processes. A method for the magnetic separation (MS) of Legionella pneumophila by superparamagnetic iron oxide nanoparticles is developed, which represents the basis for future bacteria detection kits. The focus lies on the separation process and the simplicity of using magnetic nanomaterials. Iron oxide nanoparticles are functionalized with epoxy groups and Legionella-specific antibodies are immobilized. The resulting complexes are characterized with infrared spectroscopy and tested for the specific separation and enrichment of the selected microorganisms. The cell-particle complexes can be isolated in a magnetic field and detected with conventional methods such as fluorescence detection. A nonspecific enrichment of bacteria is also possible by using bare iron oxide nanoparticles (BIONs), which we used as a reference to the nanoparticles with immobilized antibodies. Furthermore, the immunomagnetic separation can be applied for the detection of multiple other microorganisms and thus might pave the way for simpler bacterial diagnosis.

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Valciņa, Pūle, Mališevs, Trofimova, Makarova, Konvisers, Bērziņš e Krūmiņa. "Co-Occurrence of Free-Living Amoeba and Legionella in Drinking Water Supply Systems". Medicina 55, n. 8 (15 agosto 2019): 492. http://dx.doi.org/10.3390/medicina55080492.

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Background and Objectives: Legionella is one of the most important water-related pathogens. Inside the water supply systems and the biofilms, Legionella interact with other bacteria and free-living amoeba (FLA). Several amoebas may serve as hosts for bacteria in aquatic systems. This study aimed to investigate the co-occurrence of Legionella spp. and FLA in drinking water supply systems. Materials and Methods: A total of 268 water samples were collected from apartment buildings, hotels, and public buildings. Detection of Legionella spp. was performed in accordance with ISO 11731:2017 standard. Three different polymerase chain reaction (PCR) protocols were used to identify FLA. Results: Occurrence of Legionella varied from an average of 12.5% in cold water samples with the most frequent occurrence observed in hot water, in areas receiving untreated groundwater, where 54.0% of the samples were Legionella positive. The occurrence of FLA was significantly higher. On average, 77.2% of samples contained at least one genus of FLA and, depending on the type of sample, the occurrence of FLA could reach 95%. In the samples collected during the study, Legionella was always isolated along with FLA, no samples containing Legionella in the absence of FLA were observed. Conclusions: The data obtained in our study can help to focus on the extensive distribution, close interaction, and long-term persistence of Legionella and FLA. Lack of Legionella risk management plans and control procedures may promote further spread of Legionella in water supply systems. In addition, the high incidence of Legionella-related FLA suggests that traditional monitoring methods may not be sufficient for Legionella control.

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Congestrì, Francesco, Elisabetta Crepaldi, Marina Gagliardi, Maria Federica Pedna e Vittorio Sambri. "Comparative Evaluation of the Novel bioNexia Legionella Test with the BinaxNOW Legionella Card Assay and the Sofia Legionella FIA Assay for Detection of Legionella pneumophila (Serogroup 1) Antigen in Urine Samples". Journal of Clinical Microbiology 54, n. 4 (10 febbraio 2016): 1164–66. http://dx.doi.org/10.1128/jcm.03340-15.

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A new immunochromatographic test (bioNexiaLegionella; bioMérieux) for the detection ofLegionella pneumophilaurinary antigen was evaluated in 255 urine samples. The results were compared with those obtained by the BinaxNOW and SofiaLegionellatests. The novel test compared well with those currently in use.

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35

Ezenarro, Josune J., Noemí Párraga-Niño, Miquel Sabrià, Fancisco Javier Del Campo, Francesc-Xavier Muñoz-Pascual, Jordi Mas e Naroa Uria. "Rapid Detection of Legionella pneumophila in Drinking Water, Based on Filter Immunoassay and Chronoamperometric Measurement". Biosensors 10, n. 9 (20 agosto 2020): 102. http://dx.doi.org/10.3390/bios10090102.

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Abstract (sommario):

Legionella is a pathogenic bacterium, ubiquitous in freshwater environments and able to colonise man-made water systems from which it can be transmitted to humans during outbreaks. The prevention of such outbreaks requires a fast, low cost, automated and often portable detection system. In this work, we present a combination of sample concentration, immunoassay detection, and measurement by chronoamperometry. A nitrocellulose microfiltration membrane is used as support for both the water sample concentration and the Legionella immunodetection. The horseradish peroxidase enzymatic label of the antibodies permits using the redox substrate 3,3′,5,5′-Tetramethylbenzidine to generate current changes proportional to the bacterial concentration present in drinking water. Carbon screen-printed electrodes are employed in the chronoamperometric measurements. Our system reduces the detection time: from the 10 days required by the conventional culture-based methods, to 2–3 h, which could be crucial to avoid outbreaks. Additionally, the system shows a linear response (R2 value of 0.99), being able to detect a range of Legionella concentrations between 101 and 104 cfu·mL−1 with a detection limit (LoD) of 4 cfu·mL−1.

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36

Diederen, Bram M. W., Caroline M. A. de Jong, Ingrid Aarts, Marcel F. Peeters e Anneke van der Zee. "Molecular evidence for the ubiquitous presence of Legionella species in Dutch tap water installations". Journal of Water and Health 5, n. 3 (1 marzo 2007): 375–83. http://dx.doi.org/10.2166/wh.2007.033.

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Our aim was to investigate the occurrence and identity of Legionella spp. in Dutch tap water installations using culture, real-time PCR and sequence analysis. The PCR assays used were a 16S rRNA gene based PCR with both a Legionella species specific probe and a L. pneumophila specific probe and a L. pneumophila-specific PCR based on the sequence of the mip gene. A total of 357 water samples from 250 locations in The Netherlands was investigated. The detection rates of Legionella spp. were 2,2% (8 of 357) by culture, and 87,1% (311 of 357) by PCR. The majority of samples was found to contain Legionella species other than L. pneumophila. These comprised of Legionella Like Amoebal Pathogens (LLAPs), L. busanensis, L. worsliensis and others. Fourteen (3,9%) samples were positive for L. pneumophila by either culture, 16S rRNA based PCR and/or mip based PCR. It is apparent from this study that Legionella spp. DNA is ubiquitous in Dutch potable water samples. Our findings further suggest that LLAPs and viable but nonculturable (VBNC) Legionella represent a large proportion of the population in man-made environments.

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37

Jinadatha, Chetan, Eileen M. Stock, Steve E. Miller e William F. McCoy. "Environmental Validation of Legionella Control in a VHA Facility Water System". Infection Control & Hospital Epidemiology 39, n. 3 (5 febbraio 2018): 259–66. http://dx.doi.org/10.1017/ice.2017.318.

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OBJECTIVESWe conducted this study to determine what sample volume, concentration, and limit of detection (LOD) are adequate for environmental validation of Legionella control. We also sought to determine whether time required to obtain culture results can be reduced compared to spread-plate culture method. We also assessed whether polymerase chain reaction (PCR) and in-field total heterotrophic aerobic bacteria (THAB) counts are reliable indicators of Legionella in water samples from buildings.DESIGNComparative Legionella screening and diagnostics study for environmental validation of a healthcare building water system.SETTINGVeterans Health Administration (VHA) facility water system in central Texas.METHODSWe analyzed 50 water samples (26 hot, 24 cold) from 40 sinks and 10 showers using spread-plate cultures (International Standards Organization [ISO] 11731) on samples shipped overnight to the analytical lab. In-field, on-site cultures were obtained using the PVT (Phigenics Validation Test) culture dipslide-format sampler. A PCR assay for genus-level Legionella was performed on every sample.RESULTSNo practical differences regardless of sample volume filtered were observed. Larger sample volumes yielded more detections of Legionella. No statistically significant differences at the 1 colony-forming unit (CFU)/mL or 10 CFU/mL LOD were observed. Approximately 75% less time was required when cultures were started in the field. The PCR results provided an early warning, which was confirmed by spread-plate cultures. The THAB results did not correlate with Legionella status.CONCLUSIONSFor environmental validation at this facility, we confirmed that (1) 100 mL sample volumes were adequate, (2) 10× concentrations were adequate, (3) 10 CFU/mL LOD was adequate, (4) in-field cultures reliably reduced time to get results by 75%, (5) PCR provided a reliable early warning, and (6) THAB was not predictive of Legionella results.Infect Control Hosp Epidemiol 2018;39:259–266

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38

Garrison, Laurel E., Kristin M. S. Shaw, Jeffrey T. McCollum, Carol Dexter, Paula M. Snippes Vagnone, Jamie H. Thompson, Gregory Giambrone et al. "On-Site Availability of Legionella Testing in Acute Care Hospitals, United States". Infection Control & Hospital Epidemiology 35, n. 7 (luglio 2014): 898–900. http://dx.doi.org/10.1086/676871.

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Abstract (sommario):

We surveyed 399 US acute care hospitals regarding availability of on-site Legionella testing; 300 (75.2%) did not offer Legionella testing on site. Availability varied according to hospital size and geographic location. On-site access to testing may improve detection of Legionnaires disease and inform patient management and prevention efforts.Infect Control Hosp Epidemiol 2014;35(7):898–900

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39

Jahan, Rownak, Shirin Tarafder, Ahmed Abu Saleh e Ruhul Amin Miah. "Identification of Legionella from clinically diagnosed pneumonia patients and environmental samples". Bangladesh Medical Research Council Bulletin 41, n. 1 (3 novembre 2016): 24–28. http://dx.doi.org/10.3329/bmrcb.v41i1.30230.

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Abstract (sommario):

Legionnaires disease is a multisystem disease with life-threatening acute and severe form of pneumonia which is responsible for 2-9% pneumonia with high mortality. Eighty six respiratory tract samples and urine were collected from clinically diagnosed pneumonia patients and 12 water samples were collected from different environment. Identification of Legionella was done by culture and Polymerase Chain Reaction (PCR) of respiratory tract samples and environmental samples and Legionella Antigen (Ag) in urine was detected by Immunochromatographic test (ICT). Legionella was identified from 4 (4.65%) clinically diagnosed pneumonia patients of which 1 (1.16%) case was culture positive, 1 (1.16%) case was urine ICT positive and PCR was positive in all four cases. Of the 12 water samples tested, 4 (33.33%) samples were Legionella positive by PCR but culture results of these samples were negative. Identification of Legionella should be done by PCR in parallel with culture and urine ICT. Detection of Legionella in environmental samples is also needed to explore possible links between the water sources and disease transmission in population.

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Alexandropoulou, Ioanna, Theodoros Parasidis, Theocharis Konstantinidis, Maria Panopoulou e Theodoros Constantinidis. "A Proactive Environmental Approach for Preventing Legionellosis in Infants: Water Sampling and Antibiotic Resistance Monitoring, a 3-Years Survey Program". Healthcare 7, n. 1 (8 marzo 2019): 39. http://dx.doi.org/10.3390/healthcare7010039.

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Abstract (sommario):

A proactive environmental monitoring program was conducted to determine the risk and prevent nosocomial waterborne infections of Legionella spp. in infants. Sink taps in a neonatal intensive care unit (NICU) and two obstetric clinics were monitored for Legionella spp. A total of 59 water samples were collected during a 3-year period and 20 of them were found colonized with Legionella pneumophila. Standard culture, molecular, and latex agglutination methods were used for the detection and identification of Legionella bacteria. Hospital personnel also proceeded with remedial actions (hyperchlorination and thermal shock treatment) in the event of colonization. The minimal inhibitory concentration (MIC) values of erythromycin, ciprofloxacin was determined for Legionella isolates using the e-test method. Our data indicate that the majority of neonatal sink-taps were colonized at least once during the study with Legionella spp. Among 20 isolates, 5 were considered as low-level resistant, 3 in erythromycin and 2 in ciprofloxacin, while no resistant strains were detected. Environmental surveillance in neonatal and obstetric units is suggested to prevent waterborne infections, and thus to reduce the risk of neonatal nosocomial infections.

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41

Diederen, Bram M. W., Caroline M. A. de Jong, Faïçal Marmouk, Jan A. J. W. Kluytmans, Marcel F. Peeters e Anneke Van der Zee. "Evaluation of real-time PCR for the early detection of Legionella pneumophila DNA in serum samples". Journal of Medical Microbiology 56, n. 1 (1 gennaio 2007): 94–101. http://dx.doi.org/10.1099/jmm.0.46714-0.

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Abstract (sommario):

Legionella pneumonia can be difficult to diagnose. Existing laboratory tests all have shortcomings, especially in the ability to diagnose Legionnaires' disease (LD) at an early stage of the disease in a specimen that is readily obtainable. The aim of this study was to assess the performance of PCR as a rapid diagnostic method and to compare the results of different PCR assays of serum samples from patients with LD. Samples included 151 serum samples from 68 patients with proven LD and 60 serum samples from 36 patients with respiratory tract infections other than Legionella. PCR assays were based on the 5S rRNA gene, 16S rRNA gene and the mip gene. The samples from patients with infections caused by pathogens other than Legionella all tested negative in PCR. Among the patients with proven LD 54.4 % (37/68) tested positive in 5S rRNA PCR, 52.9 % (36/68) in mip gene PCR and 30.9 % (21/68) in 16S rRNA PCR in the first available serum sample. The association between threshold cycle value in 5S PCR positive serum samples (n=49) and C-reactive protein value was determined, and showed a strong negative correlation (Pearson correlation coefficient r=−0.63, P<0.0001). In addition to existing tests for the diagnosis of LD, detection of Legionella DNA in serum could be a useful tool for early diagnosis of LD caused by any Legionella species and serogroup, and has the potential to provide a diagnosis in a time frame that could affect initial infection management.

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42

Theaker, J. M., J. O. Tobin, S. E. Jones, P. Kirkpatrick, M. I. Vina e K. A. Fleming. "Immunohistological detection of Legionella pneumophila in lung sections." Journal of Clinical Pathology 40, n. 2 (1 febbraio 1987): 143–46. http://dx.doi.org/10.1136/jcp.40.2.143.

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43

Boyd, J., e E. Mcwilliams. "Immunohistological detection of Legionella pneumophila in lung sections". Journal of Clinical Pathology 40, n. 7 (1 luglio 1987): 815. http://dx.doi.org/10.1136/jcp.40.7.815-a.

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44

Iwamoto, M., H. Koga, S. Kohno, M. Kaku e K. Hara. "Detection of Legionella species by polymerase chain reaction". Serodiagnosis and Immunotherapy in Infectious Disease 7, n. 3 (settembre 1995): 99–103. http://dx.doi.org/10.1016/0888-0786(95)97891-8.

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45

Nakamura, Akihiro, Saori Fukuda, Mari Kusuki, Hideo Watari, Satoshi Shimura, Keigo Kimura, Isao Nishi e Masaru Komatsu. "Evaluation of five Legionella urinary antigen detection kits including new Ribotest Legionella for simultaneous detection of ribosomal protein L7/L12". Journal of Infection and Chemotherapy 27, n. 10 (ottobre 2021): 1533–35. http://dx.doi.org/10.1016/j.jiac.2021.05.019.

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46

Leoni, E., G. De Luca, P. P. Legnani, R. Sacchetti, S. Stampi e F. Zanetti. "Legionella waterline colonization: detection of Legionella species in domestic, hotel and hospital hot water systems". Journal of Applied Microbiology 98, n. 2 (febbraio 2005): 373–79. http://dx.doi.org/10.1111/j.1365-2672.2004.02458.x.

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47

Reuter, Cornelia, Nicole Slesiona, Stefanie Hentschel, Oliver Aehlig, Antje Breitenstein, Andrea Csáki, Thomas Henkel e Wolfgang Fritzsche. "Loop-mediated amplification as promising on-site detection approach for Legionella pneumophila and Legionella spp." Applied Microbiology and Biotechnology 104, n. 1 (12 dicembre 2019): 405–15. http://dx.doi.org/10.1007/s00253-019-10286-3.

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48

Bruin, J. P., M. F. Peeters, E. P. F. IJzerman e B. M. W. Diederen. "Evaluation of Legionella V-TesT for the detection of Legionella pneumophila antigen in urine samples". European Journal of Clinical Microbiology & Infectious Diseases 29, n. 7 (29 aprile 2010): 899–900. http://dx.doi.org/10.1007/s10096-010-0932-0.

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49

Declerck, Priscilla, Jonas Behets, Elke Lammertyn, Ilya Lebeau, Jozef Anné e Frans Ollevier. "Detection and quantification ofLegionella pneumophilain water samples using competitive PCR". Canadian Journal of Microbiology 52, n. 6 (1 giugno 2006): 584–90. http://dx.doi.org/10.1139/w05-156.

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Abstract (sommario):

The presence of high levels of Legionella pneumophila in man-made aquatic systems correlates with the incidence of nosocomial Legionnaires' disease. This requires a rapid, reliable, and sensitive quantification of L. pneumophila concentrations in suspected water systems. In this research, a homologous competitor was developed and evaluated in a L. pneumophila competitive polymerase chain reaction (cPCR) to quantify this human pathogen in a quick, cost-effective, and reliable way. Accuracy of cPCR was evaluated by analyzing cooling tower and tap water samples spiked with known concentrations of L. pneumophila bacteria, in parallel with the standard culture method. Legionella pneumophila amounts detected and calculated from cPCR and culture correlated very well: r = 0.998, P = 0.002 for tap water and r = 0.990, P = 0.009 for cooling tower water. Nevertheless, for both kinds of water samples, mean numbers of L. pneumophila calculated from cPCR results were always higher than those obtained by culture. This study makes it clear that the rapid, sensitive, and cost-effective L. pneumophila cPCR is a promising alternative to the standard time-consuming culture method and expensive real-time PCR to enumerate L. pneumophila bacteria in environmental water samples.Key words: Legionella pneumophila, competitive PCR, cost-effective, cooling tower water, tap water, sensitive detection.

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Coniglio, Maria, Margherita Ferrante e Mohamed Yassin. "Preventing Healthcare-Associated Legionellosis: Results after 3 Years of Continuous Disinfection of Hot Water with Monochloramine and an Effective Water Safety Plan". International Journal of Environmental Research and Public Health 15, n. 8 (27 luglio 2018): 1594. http://dx.doi.org/10.3390/ijerph15081594.

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The purpose of this study is to report the experience of the implementation and application of a 3-year Water Safety Plan (WSP) together with the secondary disinfection of water by monochloramine to control and prevent healthcare-associated legionellosis in an Italian hospital strongly colonized by Legionella. Risk assessment was carried out by the WSP team. The main critical control points focused on in developing the WSP for the control of Legionella was the water distribution system. A sampling plan for the detection of Legionella was implemented. A widespread contamination of the hot water distribution system by L. pneumophila sg5 was found. Results after 3 years of the continuous disinfection of hot water with monochloramine indicate the eradication of Legionella. The implementation and application of a WSP in a hospital, together with the disinfection of the water distribution system with monochloramine, can be effective in controlling the growth of Legionella and in preventing nosocomial legionellosis.

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