Novel PCR-Probe Assay for Detection of and Discrimination between
 Legionella pneumophila
 and Other
 Legionella
 Species in Clinical Samples

Zee, Anneke van der; Verbakel, Harold; Jong, Caroline de; Pot, Raymond G.J.; Bergmans, Anneke; Peeters, Marcel; Schneeberger, Peter M.; Schellekens, J. F. P.

doi:10.1128/jcm.40.3.1124-1127.2002

Cited by 30 publications

(18 citation statements)

References 5 publications

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“…from a respiratory secretion sample, detection of Legionella spp. in a respiratory secretion sample using a 16S rRNA gene PCR (16), or seroconversion to positivity for specific IgM and/or IgG antibodies (EIA; Virion/Serion GmbH, Würzburg, Germany). The positive laboratory tests of the 85 samples from LD cases were as follows: urine antigen only, 49/85 (57.6%); urine antigen and culture, 4/85 (4.7%); urine antigen and PCR, 9/85 (10.6%); urine antigen and serology, 1/85 (1.2%); urine antigen, culture, and PCR, 17/85 (20.0%); culture and PCR, 3/85 (3.5%); and culture only, 2/85 (2.4%).…”

Section: Methodsmentioning

confidence: 99%

Evaluation of the bioNexia Legionella Test, Including Impact of Incubation Time Extension, for Detection of Legionella pneumophila Serogroup 1 Antigen in Urine

et al. 2017

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In this study, we compared the bioNexia test (bioMérieux, Marcy-l'Étoile, France), a new immunochromatographic assay for the detection of Legionella pneumophila serogroup 1 in urine, with the BinaxNOW urinary antigen test (Alere, Waltham, Massachusetts, USA). After 15 min of incubation (in accordance with the manufacturers' instructions), the sensitivities and specificities were, respectively, 76.5% and 97.2% for the bioNexia test and 87.1% and 100% for the BinaxNOW test. After a prolonged incubation time of 60 min, the sensitivities and specificities increased to, respectively, 89.4% and 97.2% for the bioNexia test and 91.8% and 100% for the BinaxNOW test. When the tests were read after 15 min, the concentration of discrepant urine samples increased the sensitivities to 94.1% for both tests. In conclusion, we found that although the bioNexia test showed lower sensitivity for the detection of L. pneumophila antigen in nonconcentrated urine compared to the BinaxNOW test, a prolonged incubation time as well as the use of concentrated samples showed comparable sensitivities for both tests.KEYWORDS Legionella, Legionnaires' disease, urinary antigen test L egionnaires' disease (LD) is a severe pneumonia caused by Legionella spp., a Gram-negative bacillus found in many environments, including (man-made) aquatic systems and soil. Legionella spp. are responsible for 2 to 15% of all communityacquired pneumonias (1, 2). More than 90% of LD cases are caused by Legionella pneumophila, and 70 to 80% of these belong to serogroup type 1 (3, 4).In both Europe and the United States, the detection of Legionella antigen in urine is the most requested laboratory test for diagnosing legionnaires' disease (4). Most frequently used are immunochromatographic tests (ICTs). These tests are easy to use and are known for their relatively high sensitivity (70 to 90%) and specificity (95 to 100%), especially for L. pneumophila serogroup 1 (5-7). In addition, concentration of urine samples can increase sensitivity without affecting the specificity (3), although this preanalytical process requires some additional efforts. Heat treatment may be used to remove false-positive results caused by interfering antibodies (8). The aim of this study was to evaluate the bioNexia test (bioMérieux, Marcy-l'Étoile, France), a newly developed ICT for the qualitative detection of L. pneumophila serogroup 1 antigen in urine. When L. pneumophila serogroup 1 antigen is present in the sample, it binds to an anti-Legionella antibody that is conjugated to purple colloidal gold particles. The antigenantibody gold particle complex migrates up the membrane to the test line by capillary action. There it binds to the second anti-Legionella antibody present in the test line region. A purple-colored line is formed in the test line region. We compared the

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Section: Methodsmentioning

confidence: 99%

Evaluation of the bioNexia Legionella Test, Including Impact of Incubation Time Extension, for Detection of Legionella pneumophila Serogroup 1 Antigen in Urine

et al. 2017

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“…A PCR ELISA was performed using reagents from the Roche Diagnostic PCR ELISA kit and a Legionella-specific biotin-labelled 16S rDNA Legionella genusspecific probe (59-GGT TGC GCT CGT TAC G-39). Any positives were then retested with a Legionella pneumophila-specific biotinlabelled 16S rRNA gene probe (59-ATG TGA TGG TGG GGA CTC T-39) (van der Zee et al, 2002;Lindsay et al, 2006).…”

Section: Methodsmentioning

confidence: 99%

Legionella longbeachae serogroup 1 infections linked to potting compost

Lindsay

Brown

Brown³

et al. 2012

Journal of Medical Microbiology

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“…In recent years, different PCR-based methods (based on 16S rRNA, 5S rRNA and 23S rRNA genes and on the mip gene encoding the macrophage infectivity potentiator gene of L. pneumophila) for detection and quantification of Legionella in water samples have been described and can moderate the main drawbacks of culture-based methods [24][25][26][27][28]. The development of more rapid, culture-independent methods able to discriminating between live and dead cells is very important for assessing Legionella infection risks and preventing legionellosis [23].…”

Section: Introductionmentioning

confidence: 99%

Detection of Legionella spp. in Natural and Man-made Water Systems Using Standard Guidelines

Borges¹,

Simes²,

Martínez‐Murcia³

et al. 2012

MICROBIOLOGY

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Infections caused by Legionella spp. are considered at the present time, an emerging public health problem and are linked to high rates of mortality and morbidity, if not properly treated. In this study were analyzed 54 samples of water from 8 counties at Northern Portugal, with the aim of obtaining a collection of strains of the genus Legionella and to characterize them genetically and phenotypically. Another objective of this study was to evaluate the effectiveness of the technique of cultivation, a standard method according to International Organization for Standardization ISO 11731:1998, for detection and enumeration of species of Legionella. For laboratory processing, after the filtration of samples (1 L), the filtrate was resuspended in sterile distilled water (5 ml). Heat treatment for selective inhibition of non-Legionella bacteria was performed. Subsequently, 100 μl of the suspension was spread in GVPC selective agar medium, and incubated (7 to 10 days) at 37 ℃. Colonies that were morphologically characteristic of the genus were sub-cultured onto BCYE agar and blood agar for verification. According to the procedure recommended by the standard method, only the colonies which grew in BCYE agar and not on blood agar were considered as suspected Legionella strains. The identification of these initially selected colonies was performed by sequencing the 16S rRNA gene, which revealed that none of the isolates were identified as belonging to the genus Legionella. However, through the ISO 11731:1998 they were interpreted as positive, corresponding therefore to "false-positive" results. The methods used in this study allowed the isolation of a number of isolates (40), which form an independent group of all genus of the family Chitinophagaceae outlined so far, and that by their phylogenetic distance might be a genus not yet described and therefore a new species. The results obtained, highlighted the importance of using culture and genetic methods in parallel for the proper identification of microorganisms.

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Novel PCR-Probe Assay for Detection of and Discrimination between Legionella pneumophila and Other Legionella Species in Clinical Samples

Cited by 30 publications

References 5 publications

Evaluation of the bioNexia Legionella Test, Including Impact of Incubation Time Extension, for Detection of Legionella pneumophila Serogroup 1 Antigen in Urine

Evaluation of the bioNexia Legionella Test, Including Impact of Incubation Time Extension, for Detection of Legionella pneumophila Serogroup 1 Antigen in Urine

Legionella longbeachae serogroup 1 infections linked to potting compost

Detection of <i>Legionella</i> spp. in Natural and Man-made Water Systems Using Standard Guidelines

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