Two-Step Scheme for Rapid Identification and Differentiation of
            <i>Legionella pneumophila</i>
            and Non-
            <i>Legionella pneumophila</i>
            Species

Zhan, Xiao-Yong; Li, Lianqing; Hu, Chaohui; Zhu, Qing-Yi

doi:10.1128/jcm.01778-09

Cited by 11 publications

(5 citation statements)

References 23 publications

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“…In this study, a nested PCR was utilized to define Legionella positive water samples. The two paired primers showed high specialty for Legionella which had been described by Cloud JL et al (Cloud et al, 2000) and our research group (Zhan et al, 2010). From these seventy-six water samples, forty-two were Legionella positive according to the PCR results.…”

Section: Polymerase Chain Reaction (Pcr) Test For Water Samplesmentioning

confidence: 84%

Comparative study on sampling methods for monitoring Legionella species in environmental water

Zhan¹,

Hu²,

Zhu³

2014

Afr. J. Microbiol. Res.

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Infections caused by Legionella spp. are considered an emerging public health problem and are linked to high rates of mortality and morbidity, if not properly treated. This study analyzes 76 samples of water in Guangzhou with the aim of obtaining a more efficient means based on culture to isolate the genus Legionella. A comparative assessment of techniques recommended was performed on monitoring Legionella spp. in environmental water samples. Fourteen (14) sampling methods included in four categories based on concentration (filtration or centrifugation), acid buffer treatment (no acid treatment, treatment for 10 min), thermal treatment (no thermal treatment or treated for 30 min in 50°C) and heat enrich the samples in 37°C for 72 h were tested to identify their sensitivity. The four categories methodologies were assembled by different procedures and 14 methods were obtained to compare the efficiency for isolating Legionella in 76 water samples and in which Legionella were isolated from 42 water samples through these 14 methods. Merit and demerit of each single method was evaluated. Results suggest that a single category methodology for water sampling was insufficient to isolate Legionella spp. from water samples. Some Legionella positive water samples were missed through a single category methodology which made the results mistranslation. PSF category methods in this study were recommended methods and they could isolate Legionella species from 78.6% Legionella positive water samples; and if time was enough, PSF category methods combined with ERCT category methods to isolate Legionella from water samples are strongly recommended. Legionella were isolated from 41 water samples through these two category methods, it only missed 2.4% positive water sample. The results obtained, highlighted the importance of improving culture method for the identification of Legionella in water samples.

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Section: Polymerase Chain Reaction (Pcr) Test For Water Samplesmentioning

confidence: 84%

Comparative study on sampling methods for monitoring Legionella species in environmental water

Zhan¹,

Hu²,

Zhu³

2014

Afr. J. Microbiol. Res.

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“…PCR analysis and restriction enzyme analysis (REA) : PCR detection of the 16S rRNA gene and REA were used for identification and confirmation of L. pneumophila standard strains and environmental isolates 9 . The restriction enzyme Taa I (HpyCH4III endonuclease, Thermo-Scientific Ltd., USA) was used for the differentiation of L. pneumophila and non- pneumophila Legionella species.…”

Section: Methodsmentioning

confidence: 99%

Multiplex polymerase chain reaction of genetic markers for detection of potentially pathogenic environmental Legionella pneumophila isolates

Valavane

Chaudhry

Malhotra

2017

Indian Journal of Medical Research

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Background & objectives: Genomic constitution of the bacterium Legionella pneumophila plays an important role in providing them a pathogenic potential. Here, we report the standardization and application of multiplex polymerase chain reaction (PCR) for the detection of molecular markers of pathogenic potential in L. pneumophila in hospital environment. Methods: Culture of the standard strains of L. pneumophila was performed in buffered charcoal-yeast extract agar with L-cysteine at pH 6.9. Primers were designed for multiplex PCR, and standardization for the detection of five markers annotated to L. pneumophila plasmid pLPP (11A2), lipopolysaccharide synthesis (19H4), CMP-N-acetylneuraminic acid synthetase (10B12), conjugative coupling factor (24B1) and hypothetical protein (8D6) was done. A total of 195 water samples and 200 swabs were collected from the hospital environment. The bacterium was isolated from the hospital environment by culture and confirmed by 16S rRNA gene PCR and restriction enzyme analysis. A total of 45 L. pneumophila isolates were studied using the standardized multiplex PCR. Results: The PCR was sensitive to detect 0.1 ng/μl DNA and specific for the two standard strains used in the study. Of the 45 hospital isolates tested, 11 isolates had four markers, 12 isolates had three markers, 10 isolates had two markers, nine isolates had one marker and three isolates had none of the markers. None of the isolates had all the five markers. Interpretation & conclusions: The findings of this study showed the presence of gene markers of pathogenic potential of the bacterium L. pneumophila. However, the genomic constitution of the environmental isolates should be correlated with clinical isolates to prove their pathogenic potential. Rapid diagnostic methods such as multiplex PCR reported here, for elucidating gene markers, could help in future epidemiological studies of bacterium L. pneumophila.

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“…We also utilized a two-step assay derived method which we have previously report as a rapid and accurate means to detect and differentiate L. pneumophila and non-L. pneumophila to detect the Legionella from patient's sputum specimen. Zhan et al (2010) has reported that this two-step assay utilize 5 base sequence differences in the 16s rRNA gene between L. pneumophlia and non-L. pneumophlia to differentiate Legionella genus to different species (Zhan et al, 2010). This two-step assay include a genus specific PCR assay and a species specific enzymatic digestion analysis assay (Zhan et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

“…PCR assay was previously described in our laboratory as a conventional method (Zhan et al, 2010). Genusspecific PCR primers amplified a 386-bp portion of the 16S rRNA gene (base 451 to base 837) of Legionella.…”

Section: Pcr-enzymatic Digestion Analysismentioning

confidence: 99%

A case of malignant melanoma patient complicating pneumonia of Legionella pneumophila Sg 1 infection and means to detect Legionella from the patient

Hui¹,

Rong²,

Ming³

et al. 2013

Afr. J. Microbiol. Res.

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Legionella pneumonia is mainly caused by the inhalation of Legionella contaminated aerosol, aspiration or wound infection. Patients undergoing a severe immunosuppressive therapy such as organ transplant or cancer therapy are particularly at risk of infection. This study reported a case of Legionella pneumonia caused by Legionella pneumophila in a man with malignant melanoma. The method for diagnosing Legionella pneumonia by standard culture method was elaborate. A rapid diagnostic method derived from a two-step scheme assay with 99.42% accuracy for identification and differentiation of L. pneumophila and non-L. pneumophila in environmental Legionella isolates were also utilized in this study to evaluate whether this method was available for diagnosing Legionella infection. To confirm the diagnosis result of this rapid diagnostic method, sequencing of the bacteria in patient's sputum partial gene was also carried out. The diagnosis result of this rapid diagnostic method was consistent with the culture method which indicated that it was effective in diagnosing L. pneumophila infection.

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Two-Step Scheme for Rapid Identification and Differentiation of Legionella pneumophila and Non- Legionella pneumophila Species

Cited by 11 publications

References 23 publications

Comparative study on sampling methods for monitoring Legionella species in environmental water

Comparative study on sampling methods for monitoring Legionella species in environmental water

Multiplex polymerase chain reaction of genetic markers for detection of potentially pathogenic environmental Legionella pneumophila isolates

A case of malignant melanoma patient complicating pneumonia of Legionella pneumophila Sg 1 infection and means to detect Legionella from the patient

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