Detection of
            <i>Legionella</i>
            Species in Respiratory Specimens Using PCR with Sequencing Confirmation

Cloud, Joann L.; Carroll, Karen C.; Pixton, P.; Erali, Maria; Hillyard, David R.

doi:10.1128/jcm.38.5.1709-1712.2000

Cited by 144 publications

(50 citation statements)

References 12 publications

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“…Oligonucleotide probes used for FISH detection Protocol PCR protocol was performed using the protocol ofCloud et al (2000) with modifications. Briefly, 5 ll of extracted DNA was used in a 50-ll reaction mixture.…”

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confidence: 99%

Detection of Legionella spp. by fluorescent in situ hybridization in dental unit waterlines

et al. 2006

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Aims: To confirm the presence of viable Legionella spp. in dental unit waterlines (DUWL) using fluorescent in situ hybridization (FISH) and compare this method with culture approach and also to validate the utility of an enrichment to increase FISH sensitivity. Methods and Results: Water samples from 40 dental units were analysed. Three different techniques for detecting Legionella spp. were compared: (i) culture approach, (ii) direct FISH and (iii) FISH with a previous R2A medium enrichment (R2A/FISH). The FISH detection was confirmed by PCR. The use of the direct FISH does not improve significantly the detection of legionellae when compared with the culture. On the contrary, when R2A/FISH was performed, sensitivity was, respectively, two‐ and threefold higher than that with the direct FISH and culture approach. Using R2A/FISH, 63% of water samples analysed showed a contamination by legionellae. Conclusions: Legionellae detection by direct FISH and R2A/FISH in dental unit water is possible but is more rapid and more sensitive (R2A/FISH) than the culture approach. Significance and Impact of the Study: R2A/FISH showed that several pathogens present in DUWL are viable but may not be culturable. Unlike PCR, R2A/FISH is designed to detect only metabolically active cells and therefore provides more pertinent information on infectious risk.

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confidence: 99%

Detection of Legionella spp. by fluorescent in situ hybridization in dental unit waterlines

et al. 2006

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“…Ces PCR sont réalisables théoriquement en moins de 24 h, sur tout type de prélèvement respiratoire mais sont encore actuellement trop peu utilisées. Elles auraient l'avantage d'identifier les cas de légionelloses à L. pneumophila sérogroupe non 1, voire à Legionella autres que L. pneumophila [43] [49,50]. L'addition dans ces différents panels respiratoires de cibles bactériennes est beaucoup plus récente et les données de sensibilité et de spécificité sont encore quasi-inexistantes.…”

Section: Apports De La Biologie Moléculaire Dans Les Infections Des Vunclassified

Description et place des techniques bactériologiques dans la prise en charge des infections pulmonaires

Dahyot

Lemée

Pestel-Caron

2017

Revue des Maladies Respiratoires

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Acute pneumonias occur in a variety of clinical settings and accurate identification of bacterial causes is extremely important. No microbiological tool is either 100 % sensitive or 100 % specific, and despite investigations, aetiology remains unanswered in more than 30 % of pneumonia. No sample may be necessary for patients treated as outpatients, non invasive respiratory specimens are preferred in hospitalised individuals (community or healthcare associated), while invasive specimens are used as second line for community acquired pneumonia (CAP) in intensive care, and in the first line where pneumonia occurs in immunosuppressed patients. Bacterial cultures have an important place, if the sample is taken before the introduction of antibiotic therapy. Some contexts may justify the use of blood cultures, testing for urinary antigens or serology. PCR is already becoming available as a daily service but the short-term future probably belongs to molecular multiplex panels capable of detecting many microorganisms within hours, especially in severe CAP resuscitation and in pneumonia in the immunosuppressed. High-throughput sequencing nucleotide techniques will soon revolutionize microbiological diagnosis in respiratory medicine, as in other areas of infectious diseases.

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“…Notably, these non-FDA cleared assays include the ones for atypical agents, such as C pneumoniae, M pneumoniae, and Legionella pneumophilia. [66][67][68][69][70][71]…”

Section: Non-fda-cleared Molecular Assays For Bacteriamentioning

confidence: 99%

What Is the Role of Newer Molecular Tests in the Management of CAP?

Gaydos

2013

Infectious Disease Clinics of North America

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Community-acquired pneumonia (CAP) accounts for major morbidity and mortality in the United States. With improved broad-spectrum antibiotics, the implementation of diagnostic studies has declined and most patients do not have an etiologic pathogen of CAP identified. To enhance the appropriate use of antiviral agents and prevent overuse of antibiotics, the successful management of CAP requires rapid and accurate diagnosis of the etiologic agent of CAP. This article provides an overview of the new rapid molecular tests for the diagnosis of influenza, other respiratory viruses, and bacteria compared with nonmolecular tests and how their use for directed therapy can enhance and improve the management of CAP.

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Detection of Legionella Species in Respiratory Specimens Using PCR with Sequencing Confirmation

Cited by 144 publications

References 12 publications

Detection of Legionella spp. by fluorescent in situ hybridization in dental unit waterlines

Detection of Legionella spp. by fluorescent in situ hybridization in dental unit waterlines

Description et place des techniques bactériologiques dans la prise en charge des infections pulmonaires

What Is the Role of Newer Molecular Tests in the Management of CAP?

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