Evaluation of Legionella real-time PCR against traditional culture for routine and public health testing of water samples

S. Collins; D. Stevenson; J. Walker; Allan Bennett

doi:10.1111/jam.13461

Evaluation of Legionella real-time PCR against traditional culture for routine and public health testing of water samples

S. Collins^*, D. Stevenson, J. Walker, Allan Bennett

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

24 Citations (Scopus)

Abstract

Aims: To evaluate the usefulness of Legionella qPCR alongside traditional culture for enumeration of Legionella from water samples as part of both routine and public health investigation testing. Methods and Results: Routine water samples (n = 2002) and samples from public health investigations (n = 215) were analysed by culture and qPCR for Legionella spp., Legionella pneumophila and L. pneumophila sg-1. A negative qPCR result was highly predictive of a negative culture result for all water systems (negative predictive values, NPV from 97·4 to 100%). Positive predictive values (PPV) were lower (0–50%). Results for qPCR were generally larger than culture with average log₁₀ differences of 1·1 for Legionella spp. and 1·2 for L. pneumophila. Alert and action levels of 1000 and 10 000 GU per litre, respectively, are proposed for Legionella qPCR for hot and cold water systems (HCWS). The use of qPCR significantly reduced the time to results for public health investigations by rapidly identifying potential sources and ruling out others, thus enabling a more rapid and efficient response. Conclusions: The high NPV of qPCR supports its use to rapidly screen out negative samples without culture. Alert and action levels for Legionella qPCR for HCWS are proposed. Quantitative PCR will be a valuable tool for both routine and public health testing. Significance and Impact of the Study: This study generated comparative data of >2000 water samples by qPCR and culture. Action and alert levels have been recommended that could enable duty holders to interpret qPCR results to facilitate timely Legionella control and public health protection.

Original language	English
Pages (from-to)	1692-1703
Number of pages	12
Journal	Journal of Applied Microbiology
Volume	122
Issue number	6
DOIs	https://doi.org/10.1111/jam.13461
Publication status	Published - Jun 2017

Bibliographical note

Funding Information:
The authors thank the PHE Food, Water and Environmental Laboratory network for access to samples and support throughout the duration of the project. Cooling tower samples were kindly provided by Assurity Consulting. Thanks are due to Andr? Charlet for statistical support, Paul McDermott, John Newbold and all agencies and local authorities involved in the public health investigations. This study was funded by Public Health England and the Health and Safety Executive and the views expressed in this publication are those of the authors and not necessarily those of Public Health England or the Health and Safety Executive.

Keywords

Legionella
Legionnaires’ disease
culture
environmental
qPCR
water samples

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1111/jam.13461

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title = "Evaluation of Legionella real-time PCR against traditional culture for routine and public health testing of water samples",

abstract = "Aims: To evaluate the usefulness of Legionella qPCR alongside traditional culture for enumeration of Legionella from water samples as part of both routine and public health investigation testing. Methods and Results: Routine water samples (n = 2002) and samples from public health investigations (n = 215) were analysed by culture and qPCR for Legionella spp., Legionella pneumophila and L. pneumophila sg-1. A negative qPCR result was highly predictive of a negative culture result for all water systems (negative predictive values, NPV from 97·4 to 100%). Positive predictive values (PPV) were lower (0–50%). Results for qPCR were generally larger than culture with average log10 differences of 1·1 for Legionella spp. and 1·2 for L. pneumophila. Alert and action levels of 1000 and 10 000 GU per litre, respectively, are proposed for Legionella qPCR for hot and cold water systems (HCWS). The use of qPCR significantly reduced the time to results for public health investigations by rapidly identifying potential sources and ruling out others, thus enabling a more rapid and efficient response. Conclusions: The high NPV of qPCR supports its use to rapidly screen out negative samples without culture. Alert and action levels for Legionella qPCR for HCWS are proposed. Quantitative PCR will be a valuable tool for both routine and public health testing. Significance and Impact of the Study: This study generated comparative data of >2000 water samples by qPCR and culture. Action and alert levels have been recommended that could enable duty holders to interpret qPCR results to facilitate timely Legionella control and public health protection.",

keywords = "Legionella, Legionnaires{\textquoteright} disease, culture, environmental, qPCR, water samples",

author = "S. Collins and D. Stevenson and J. Walker and Allan Bennett",

note = "Funding Information: The authors thank the PHE Food, Water and Environmental Laboratory network for access to samples and support throughout the duration of the project. Cooling tower samples were kindly provided by Assurity Consulting. Thanks are due to Andr? Charlet for statistical support, Paul McDermott, John Newbold and all agencies and local authorities involved in the public health investigations. This study was funded by Public Health England and the Health and Safety Executive and the views expressed in this publication are those of the authors and not necessarily those of Public Health England or the Health and Safety Executive.",

year = "2017",

month = jun,

doi = "10.1111/jam.13461",

language = "English",

volume = "122",

pages = "1692--1703",

journal = "Journal of Applied Microbiology",

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AU - Collins, S.

AU - Stevenson, D.

AU - Walker, J.

AU - Bennett, Allan

N1 - Funding Information: The authors thank the PHE Food, Water and Environmental Laboratory network for access to samples and support throughout the duration of the project. Cooling tower samples were kindly provided by Assurity Consulting. Thanks are due to Andr? Charlet for statistical support, Paul McDermott, John Newbold and all agencies and local authorities involved in the public health investigations. This study was funded by Public Health England and the Health and Safety Executive and the views expressed in this publication are those of the authors and not necessarily those of Public Health England or the Health and Safety Executive.

PY - 2017/6

Y1 - 2017/6

N2 - Aims: To evaluate the usefulness of Legionella qPCR alongside traditional culture for enumeration of Legionella from water samples as part of both routine and public health investigation testing. Methods and Results: Routine water samples (n = 2002) and samples from public health investigations (n = 215) were analysed by culture and qPCR for Legionella spp., Legionella pneumophila and L. pneumophila sg-1. A negative qPCR result was highly predictive of a negative culture result for all water systems (negative predictive values, NPV from 97·4 to 100%). Positive predictive values (PPV) were lower (0–50%). Results for qPCR were generally larger than culture with average log10 differences of 1·1 for Legionella spp. and 1·2 for L. pneumophila. Alert and action levels of 1000 and 10 000 GU per litre, respectively, are proposed for Legionella qPCR for hot and cold water systems (HCWS). The use of qPCR significantly reduced the time to results for public health investigations by rapidly identifying potential sources and ruling out others, thus enabling a more rapid and efficient response. Conclusions: The high NPV of qPCR supports its use to rapidly screen out negative samples without culture. Alert and action levels for Legionella qPCR for HCWS are proposed. Quantitative PCR will be a valuable tool for both routine and public health testing. Significance and Impact of the Study: This study generated comparative data of >2000 water samples by qPCR and culture. Action and alert levels have been recommended that could enable duty holders to interpret qPCR results to facilitate timely Legionella control and public health protection.

AB - Aims: To evaluate the usefulness of Legionella qPCR alongside traditional culture for enumeration of Legionella from water samples as part of both routine and public health investigation testing. Methods and Results: Routine water samples (n = 2002) and samples from public health investigations (n = 215) were analysed by culture and qPCR for Legionella spp., Legionella pneumophila and L. pneumophila sg-1. A negative qPCR result was highly predictive of a negative culture result for all water systems (negative predictive values, NPV from 97·4 to 100%). Positive predictive values (PPV) were lower (0–50%). Results for qPCR were generally larger than culture with average log10 differences of 1·1 for Legionella spp. and 1·2 for L. pneumophila. Alert and action levels of 1000 and 10 000 GU per litre, respectively, are proposed for Legionella qPCR for hot and cold water systems (HCWS). The use of qPCR significantly reduced the time to results for public health investigations by rapidly identifying potential sources and ruling out others, thus enabling a more rapid and efficient response. Conclusions: The high NPV of qPCR supports its use to rapidly screen out negative samples without culture. Alert and action levels for Legionella qPCR for HCWS are proposed. Quantitative PCR will be a valuable tool for both routine and public health testing. Significance and Impact of the Study: This study generated comparative data of >2000 water samples by qPCR and culture. Action and alert levels have been recommended that could enable duty holders to interpret qPCR results to facilitate timely Legionella control and public health protection.

KW - Legionella

KW - Legionnaires’ disease

KW - culture

KW - environmental

KW - qPCR

KW - water samples

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JO - Journal of Applied Microbiology

JF - Journal of Applied Microbiology

IS - 6

ER -

Evaluation of Legionella real-time PCR against traditional culture for routine and public health testing of water samples

Abstract

Bibliographical note

Keywords

UN SDGs

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