Plasma‐activated carbon nanotube‐based high sensitivity immunosensors for monitoring Legionella pneumophila by direct detection of maltose binding protein peptidoglycan‐associated lipoprotein (MBP‐PAL)

Detection of Legionella by quantitative-Polymerase Chain Reaction (qPCR) for monitoring and risk assessment

Research article Louise H Krojgaard, Karen A Krogfelt, Hans-Jorgen Albrechtsen and Soren A Uldum

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BMC Microbiology 2011, 11:254 doi:10.1186/1471-2180-11-254

Published: 21 November 2011

Background Culture and quantitative polymerase chain reaction (qPCR) assays for the detection of Legionella were compared on samples from a residential area before and after two interventions. A total of 84 samples were collected from shower hoses and taps as first flush samples and at constant temperature. Samples were grouped according to the origin of the sample, a) circulation water b) water from empty apartments c) water from shower hoses. The aims were to investigate the usefulness of qPCR compared to culture for monitoring remedial actions for elimination of Legionella bacteria and as a tool for risk assessment.

In water collected from the apartments Legionella spp were detected by qPCR in the concentration range from LOQ to 9.6*105GU/L while L. pneumophila were detected in a range from LOQ to 6.8 *105 GU/L. By culturing, the legionellae were detected in the range from below detection limit (> 10 CFU/L) to 1.6 * 106 CFU/L. In circulating water and in first flush water from shower hoses, culture and qPCR showed the same tendencies. The overall correlation between the bacteria number detected by culture and the two developed qPCR assays (L. spp and L. pneumophila) was relatively poor (r2 =0.31 for culture and Legionella spp. assay, r2 = 0.20 for culture and L. pneumophila assay).

Detection by qPCR was suitable for monitoring changes in the concentration of Legionella but the precise determination of bacteria is difficult. Risk assessment by qPCR only on samples without any background information regarding treatment, timing, etc is dubious. However, the rapid detection by qPCR of high concentrations of Legionella - especially Legionella pneumophila - is valuable as an indicator of risk, although it may be false positive compared to culture results. On the other hand, the detection of a low number of bacteria by qPCR is a strong indication for the absence of risk.

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Detection of Legionella by quantitative-Polymerase Chain Reaction (qPCR) for monitoring and risk assessment

Conclusion:Detection by qPCR was suitable for monitoring changes in the concentration of Legionella but the precise determination of bacteria is difficult. Risk assessment by qPCR only on samples without any background information regarding treatment, timing, etc is dubious. However, the rapid detection by qPCR of high concentrations of Legionella - especially Legionella pneumophila - is valuable as an indicator of risk, although it may be false positive compared to culture results. On the other hand, the detection of a low number of bacteria by qPCR is a strong indication for the absence of risk. (Source: BMC Microbiology - Latest articles)

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The immunochromatic kits Xpect(R) Legionella and BinaxNOW(R) Legionella for detection of Legionella pneumophila urinary antigen have low sensitivities for the diagnosis of Legionnaires' disease.

Detection of microorganisms in exhaled breath condensate during acute exacerbations of COPD

Conclusions: Bacterial nucleic acids can be identified in EBCs of COPD patients with exacerbations. The results obtained from EBC and sputum did not correlate well. (Source: Respirology)

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Detection of microorganisms in exhaled breath condensate during acute exacerbations of COPD

Detection of microorganisms in exhaled breath condensate during acute exacerbations of COPD - Zakharkina - Respirology - Wiley Online LibrarySkip to Main Content

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PUBLICATIONSBROWSE BY SUBJECTRESOURCESABOUT US LOGIN Enter e-mail address Enter password REMEMBER ME NOT REGISTERED ?FORGOTTEN PASSWORD ?INSTITUTIONAL LOGIN > Home > Respiratory Medicine > Respiratory Medicine > Journal Home > Accepted Articles > Abstract JOURNAL TOOLS Get New Content Alerts Get RSS feed Save to My Profile Get Sample Copy Recommend to Your Librarian JOURNAL MENU Journal Home FIND ISSUES Current IssueAll IssuesVirtual Issues FIND ARTICLES Accepted Articles GET ACCESS Subscribe / Renew FOR CONTRIBUTORS Author GuidelinesSubmit an Article ABOUT THIS JOURNAL OverviewNewsEditorial BoardContactSociety InformationAdvertisePermissions Detection of microorganisms in exhaled breath condensate during acute exacerbations of COPDTetyana Zakharkina1,2,†, A.-Rembert Koczulla1,†, Olga Mardanova1, Akira Hattesohl1, Robert Bals1,2,*DOI: 10.1111/j.1440-1843.2011.01977.x

© 2011 The Authors. Respirology © 2011 Asian Pacific Society of Respirology

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RespirologyAccepted Article (Accepted, unedited articles published online for future issues)

Additional Information

Author Information1

Department of Internal Medicine, Division for Pulmonary Diseases, University Hospital Giessen and Marburg, D-35043 Marburg, Germany

2

Department of Internal Medicine V -Pulmonology, Respiratory Intensive Care Medicine, Allergology, D-66421 Homburg, Germany

*Correspondence: Robert Bals,

*Correspondence: Prof. Dr. Dr. Robert Bals ?Department of Internal Medicine V -Pulmonology, Respiratory Intensive Care Medicine, Allergology ?D-66421 Homburg, Germany ?Tel: +49 (0)6841 16 23601 Fax: +49 (0) 6841 16 23602 ?E-mail: robert.bals@uks.eu

†

both authors contributed equally

This is an Accepted Article that has been peer-reviewed and approved for publication in the Respirology, but has yet to undergo copy-editing and proof correction. Please cite this article as an “Accepted Article”; doi: 10.1111/j.1440-1843.2011.01977.x

Publication HistoryAccepted manuscript online: 7 APR 2011 01:59AM ESTAccepted 21 February 2011 SEARCH Search Scope All contentPublication titlesIn this journalIn this issue Search String Advanced >Saved Searches > SEARCH BY CITATION Volume: Issue: Page: ARTICLE TOOLSGet PDF (194K)Save to My ProfileE-mail Link to this ArticleExport Citation for this ArticleGet Citation AlertsRequest Permissions AbstractCited By Get PDF (194K) Keywords:COPD;exacerbation;exhaled breath condensate;sputum;bacteria;virusesSummary at a Glance

Exhaled breath condensate (EBC) is a noninvasive method to assess airway biology. The aim of the present study was to evaluate EBC for the detection of microbial nucleic acids during acute exacerbations of COPD (AECOPD). Bacterial nucleic acids can be identified in EBCs of COPD patients with exacerbations, however, the results differed significantly from those of sputum.

ABSTRACT

Background and objective: One hallmark of chronic obstructive pulmonary disease (COPD) is colonization and infection of the lung. Acute exacerbations of COPD (AECOPD) are acute deteriorations of the chronic disease and are associated with a change of the pulmonary microbial balance. The collection of exhaled breath condensate (EBC) can be used to noninvasively determine markers of lung disease. The aim of the present study was to compare the results of assays based on the detection of microbial nucleic acids from exhaled breath condensate (EBC) and from spontaneous sputum in patients with AECOPD.

Methods: EBCs and sputa of 29 adults with AECOPD were obtained. Isolated DNA or RNA were used as starting material for the PCR assays to detect Staphylococcus aureus, Haemophilus influenzae, Moraxella catarrhalis, Streptococcus pneumoniae, Legionella pneumophila,Mycoplasma pneumoniae,Chlamydia pneumoniae, influenza viruses (AH 1, AH 3) and respiratory syncytial virus.

Results: Bacterial or viral nucleic acids were identified in 14 EBCs and 21 sputa from 29 patients. Results from EBC did not correlate well with those from sputum. Viral and S. pneumoniae nucleic acids were detected only in sputum, whereas L. pneumophila DNA was only found in EBC. In 3 EBCs and 10 sputa nucleic acids of more than one microorganism was detected.

Conclusions: Bacterial nucleic acids can be identified in EBCs of COPD patients with exacerbations. The results obtained from EBC and sputum did not correlate well.

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