ArticleApplied and environmental microbiology2005
Quantitative detection of Legionella pneumophila in water samples by immunomagnetic purification and real-time PCR amplification of the dotA gene.
Article in Applied and environmental microbiology, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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30 citing papers in PubMed, 110 citations in OpenAlex.
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- Spatial-temporal targeted and non-targeted surveys to assess microbiological composition of drinking water in Puerto Rico following Hurricane Maria.Water research X · 2021Article
- Comparison of the culture method with multiplex PCR for the confirmation of Legionella spp. and Legionella pneumophila.Journal of applied microbiology · 2021Article
- Quantification of Legionella pneumophila by qPCR and culture in tap water with different concentrations of residual disinfectants and heterotrophic bacteria.The Science of the total environment · 2021Article
- Rapid Detection and Differentiation ofFrontiers in chemistry · 2021Article
- Development of a DGGE method to exploreHeliyon · 2020Article
- Persistent presence of outer membrane epitopes during short- and long-term starvation of five Legionella pneumophila strains.BMC microbiology · 2018Article
- Properties of bacterial communities attached to artificial substrates in a hypereutrophic urban river.AMB Express · 2018Article
- Comparison of Database Search Methods for the Detection ofFrontiers in microbiology · 2018Article
- High Prevalence and Genetic Polymorphisms of Legionella in Natural and Man-Made Aquatic Environments in Wenzhou, China.International journal of environmental research and public health · 2017Article
- Occurrence of Legionella in wastewater treatment plants linked to wastewater characteristics.Environmental science and pollution research international · 2016Article
- Combined use of real-time PCR and nested sequence-based typing in survey of human Legionella infection.Epidemiology and infection · 2016Article
- PCR method for the rapid detection and discrimination of Legionella spp. based on the amplification of pcs, pmtA, and 16S rRNA genes.Journal of applied genetics · 2016Article
- Surveillance of parasitic Legionella in surface waters by using immunomagnetic separation and amoebae enrichment.Pathogens and global health · 2015Article
- Current and emerging Legionella diagnostics for laboratory and outbreak investigations.Clinical microbiology reviews · 2015Review
- Evaluation of immunomagnetic separation for the detection of Salmonella in surface waters by polymerase chain reaction.International journal of environmental research and public health · 2014Article
- Limitations of Using Propidium Monoazide with qPCR to Discriminate between Live and Dead Legionella in Biofilm Samples.Microbiology insights · 2014Article
- Fast immunosensing technique to detect Legionella pneumophila in different natural and anthropogenic environments: comparative and collaborative trials.BMC microbiology · 2013Article
- Distribution of Legionella pneumophila bacteria and Naegleria and Hartmannella amoebae in thermal saline baths used in balneotherapy.Parasitology research · 2013Article
- Rapid detection of total and viable Legionella pneumophila in tap water by immunomagnetic separation, double fluorescent staining and flow cytometry.Microbial biotechnology · 2012Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A new real-time PCR assay was developed and validated in combination with an immunomagnetic separation system for the quantitative determination of Legionella pneumophila in water samples. Primers that amplify simultaneously an 80-bp fragment of the dotA gene from L. pneumophila and a recombinant fragment including a specific sequence of the gyrB gene from Aeromonas hydrophila, added as an internal positive control, were used. The specificity, limit of detection, limit of quantification, repetitivity, reproducibility, and accuracy of the method were calculated, and the values obtained confirmed the applicability of the method for the quantitative detection of L. pneumophila. Moreover, the efficiency of immunomagnetic separation in the recovery of L. pneumophila from different kinds of water was evaluated. The recovery rates decreased as the water contamination increased (ranging from 59.9% for distilled water to 36% for cooling tower water), and the reproducibility also decreased in parallel to water complexity. The feasibility of the method was evaluated by cell culture and real-time PCR analysis of 60 samples in parallel. All the samples found to be positive by cell culture were also positive by real-time PCR, while only eight samples were found to be positive only by PCR. Finally, the correlation of both methods showed that the number of cells calculated by PCR was 20-fold higher than the culture values. In conclusion, the real-time PCR method combined with immunomagnetic separation provides a sensitive, specific, and accurate method for the rapid quantification of L. pneumophila in water samples. However, the recovery efficiency of immunomagnetic separation should be considered in complex samples.
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