Simultaneous detection of Legionella species and L. anisa , L. bozemanii , L. longbeachae and L. micdadei using conserved primers and multiple probes in a multiplex real-time PCR assay

Cross, Kristen E.; Mercante, Jeffrey W.; Benitez, Alvaro J.; Brown, Ellen; Diaz, Maureen H.; Winchell, Jonas M.

doi:10.1016/j.diagmicrobio.2016.03.022

Cited by 22 publications

(13 citation statements)

References 41 publications

(59 reference statements)

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“…by direct fluorescent-antibody staining using monoclonal-antibody derived reagents, (2) detection of Legionella spp. nucleic acid in respiratory secretions, lung tissue or any normally sterile site [28,29], (3) significant rise in specific antibody level to L. pneumophila other than serogroup 1 or other Legionella spp. in paired serum samples, single high level of specific antibody to L. pneumophila serogroup 1 in serum.…”

Section: Methodsmentioning

confidence: 99%

Short-term associations between Legionnaires' disease incidence and meteorological variables in Belgium, 2011–2019

et al. 2020

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The number of reported cases with Legionnaires' disease (LD) is increasing in Belgium. Previous studies have investigated the associations between LD incidence and meteorological factors, but the Belgian data remained unexplored. We investigated data collected between 2011 and 2019. Daily exposure data on temperature, relative humidity, precipitation and wind speed was obtained from the Royal Meteorological Institute for 29 weather stations. Case data were collected from the national reference centre and through mandatory notification. Daily case and exposure data were aggregated by province. We conducted a time-stratified case-crossover study. The ‘at risk’ period was defined as 10 to 2 days prior to disease onset. The corresponding days in the other study years were selected as referents. We fitted separate conditional Poisson models for each day in the ‘at risk’ period and a distributed lag non-linear model (DLNM) which fitted all data in one model. LD incidence showed a yearly peak in August and September. A total of 614 cases were included. Given seasonality, a sequence of precipitation, followed by high relative humidity and low wind speed showed a statistically significant association with the number of cases 6 to 4 days later. We discussed the advantages of DLNM in this context.

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Section: Methodsmentioning

confidence: 99%

Short-term associations between Legionnaires' disease incidence and meteorological variables in Belgium, 2011–2019

et al. 2020

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“…Raw data of Five Disease Relevant Legionella species. References [54,55] are cited in the Supplementary Materials.…”

Section: Supplementary Materialsmentioning

confidence: 99%

Water Age Effects on the Occurrence and Concentration of Legionella Species in the Distribution System, Premise Plumbing, and the Cooling Towers

Logan-Jackson

Rose

2021

Microorganisms

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In this study, droplet digital PCRTM (ddPCRTM) was used to characterize total Legionella spp. and five specific Legionella species from source (groundwater) to exposure sites (taps and cooling towers). A total of 42–10 L volume water samples were analyzed during this study: 12 from a reservoir (untreated groundwater and treated water storage tanks), 24 from two buildings (influents and taps), and six from cooling towers, all part of the same water system. The approximate water age (time in the system) for all sample locations are as follows: ~4.5, 3.4, 9.2, 20.8, and 23.2 h (h) for the groundwater to the reservoir influent, reservoir influent to the reservoir effluent, reservoir effluent to building Fa (building names are abbreviated to protect the privacy of site location), building ERC and the cooling towers, respectively. Results demonstrated that gene copies of Legionella spp. (23S rRNA) were significantly higher in the cooling towers and ERC building (p < 0.05) relative to the reservoir and building Fa (closest to reservoir). Legionella spp. (23S rRNA) were found in 100% (42/42) of water samples at concentrations ranging from 2.2 to 4.5 Log10 GC/100 mL. More specifically, L. pneumophila was found in 57% (24/42) of the water samples, followed by L. bozemanii 52% (22/42), L. longbeachae 36% (15/42), L. micdadei 23% (10/42) and L. anisa 21% (9/42) with geometric mean concentrations of 1.7, 1.7, 1.4, 1.6 and 1.7 Log10 GC/100 mL, respectively. Based on this study, it is hypothesized that water age in the distribution system and the premise-plumbing system as well as building management plays a major role in the increase of Legionella spp., (23S rRNA) and the diversity of pathogenic species found as seen in the influent, and at the taps in the ERC building—where the building water quality was most comparable to the industrial cooling towers. Other pathogenic Legionella species besides L.pneumophila are also likely amplifying in the system; thus, it is important to consider other disease relevant species in the whole water supply system—to subsequently control the growth of pathogenic Legionella in the built water environment.

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“…All primers and probes were designed and validated previously by the authors listed in the references below. Not available [69] Positive controls using genomic DNA from amoebae, A. castellani strain Neff (ATCC No. 30010) and N. fowleri (ATCC No.…”

Section: Molecular Analysismentioning

confidence: 99%

Cooccurrence of Five Pathogenic Legionella spp. and Two Free-Living Amoebae Species in a Complete Drinking Water System and Cooling Towers

Logan-Jackson

Rose

2021

Pathogens

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Pathogenic Legionella species grow optimally inside free-living amoebae to concentrations that increase risks to those who are exposed. The aim of this study was to screen a complete drinking water system and cooling towers for the occurrence of Acanthamoeba spp. and Naegleria fowleri and their cooccurrence with Legionella pneumophila, Legionella anisa, Legionella micdadei, Legionella bozemanii, and Legionella longbeachae. A total of 42 large-volume water samples, including 12 from the reservoir (water source), 24 from two buildings (influents to the buildings and exposure sites (taps)), and six cooling towers were collected and analyzed using droplet digital PCR (ddPCR). N. fowleri cooccurred with L. micdadei in 76 (32/42) of the water samples. In the building water system, the concentrations of N. fowleri and L. micdadei ranged from 1.5 to 1.6 Log10 gene copies (GC)/100 mL, but the concentrations of species increased in the cooling towers. The data obtained in this study illustrate the ecology of pathogenic Legionella species in taps and cooling towers. Investigating Legionella’s ecology in drinking and industrial waters will hopefully lead to better control of these pathogenic species in drinking water supply systems and cooling towers.

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Simultaneous detection of Legionella species and L. anisa , L. bozemanii , L. longbeachae and L. micdadei using conserved primers and multiple probes in a multiplex real-time PCR assay

Cited by 22 publications

References 41 publications

Short-term associations between Legionnaires' disease incidence and meteorological variables in Belgium, 2011–2019

Short-term associations between Legionnaires' disease incidence and meteorological variables in Belgium, 2011–2019

Water Age Effects on the Occurrence and Concentration of Legionella Species in the Distribution System, Premise Plumbing, and the Cooling Towers

Cooccurrence of Five Pathogenic Legionella spp. and Two Free-Living Amoebae Species in a Complete Drinking Water System and Cooling Towers

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