Multiplex real-time PCR assay for Legionella species

Seung Min Kim; Yoojung Jeong; Jang Wook Sohn; Min Ja Kim

doi:10.1016/j.mcp.2015.08.004

Multiplex real-time PCR assay for Legionella species

Seung Min Kim, Yoojung Jeong, Jang Wook Sohn, Min Ja Kim

Department of Biomedical Sciences

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

3 Citations (Scopus)

Abstract

Legionella pneumophila serogroup 1 (sg1) accounts for the majority of infections in humans, but other Legionella species are also associated with human disease. In this study, a new SYBR Green I-based multiplex real-time PCR assay in a single reaction was developed to allow the rapid detection and differentiation of Legionella species by targeting specific gene sequences. Candidate target genes were selected, and primer sets were designed by referring to comparative genomic hybridization data of Legionella species. The Legionella species-specific groES primer set successfully detected all 30 Legionella strains tested. The xcpX and rfbA primers specifically detected L. pneumophila sg1-15 and L. pneumophila sg1, respectively. In addition, this assay was validated by testing clinical samples and isolates. In conclusion, this novel multiplex real-time PCR assay might be a useful diagnostic tool for the rapid detection and differentiation of Legionella species in both clinical and epidemiological studies.

Original language	English
Pages (from-to)	414-419
Number of pages	6
Journal	Molecular and Cellular Probes
Volume	29
Issue number	6
DOIs	https://doi.org/10.1016/j.mcp.2015.08.004
Publication status	Published - 2015 Dec 1

Keywords

Legionella species
Legionnaires' disease
Multiplex
Real-time PCR

ASJC Scopus subject areas

Molecular Biology
Cell Biology

UN SDGs

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

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10.1016/j.mcp.2015.08.004

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title = "Multiplex real-time PCR assay for Legionella species",

abstract = "Legionella pneumophila serogroup 1 (sg1) accounts for the majority of infections in humans, but other Legionella species are also associated with human disease. In this study, a new SYBR Green I-based multiplex real-time PCR assay in a single reaction was developed to allow the rapid detection and differentiation of Legionella species by targeting specific gene sequences. Candidate target genes were selected, and primer sets were designed by referring to comparative genomic hybridization data of Legionella species. The Legionella species-specific groES primer set successfully detected all 30 Legionella strains tested. The xcpX and rfbA primers specifically detected L. pneumophila sg1-15 and L. pneumophila sg1, respectively. In addition, this assay was validated by testing clinical samples and isolates. In conclusion, this novel multiplex real-time PCR assay might be a useful diagnostic tool for the rapid detection and differentiation of Legionella species in both clinical and epidemiological studies.",

keywords = "Legionella species, Legionnaires' disease, Multiplex, Real-time PCR",

author = "Kim, {Seung Min} and Yoojung Jeong and Sohn, {Jang Wook} and Kim, {Min Ja}",

note = "Funding Information: This study was financially supported in part by Yuhan Co. (Seoul, Korea), the City of Seoul Grant # 10920 , and the National Research Foundation of Korea grant funded by the Korean government (No. K20902001448-10E0100-03010 ). Publisher Copyright: {\textcopyright} 2015 Elsevier Ltd.",

year = "2015",

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doi = "10.1016/j.mcp.2015.08.004",

language = "English",

volume = "29",

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journal = "Molecular and Cellular Probes",

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T1 - Multiplex real-time PCR assay for Legionella species

AU - Kim, Seung Min

AU - Jeong, Yoojung

AU - Sohn, Jang Wook

AU - Kim, Min Ja

N1 - Funding Information: This study was financially supported in part by Yuhan Co. (Seoul, Korea), the City of Seoul Grant # 10920 , and the National Research Foundation of Korea grant funded by the Korean government (No. K20902001448-10E0100-03010 ). Publisher Copyright: © 2015 Elsevier Ltd.

PY - 2015/12/1

Y1 - 2015/12/1

N2 - Legionella pneumophila serogroup 1 (sg1) accounts for the majority of infections in humans, but other Legionella species are also associated with human disease. In this study, a new SYBR Green I-based multiplex real-time PCR assay in a single reaction was developed to allow the rapid detection and differentiation of Legionella species by targeting specific gene sequences. Candidate target genes were selected, and primer sets were designed by referring to comparative genomic hybridization data of Legionella species. The Legionella species-specific groES primer set successfully detected all 30 Legionella strains tested. The xcpX and rfbA primers specifically detected L. pneumophila sg1-15 and L. pneumophila sg1, respectively. In addition, this assay was validated by testing clinical samples and isolates. In conclusion, this novel multiplex real-time PCR assay might be a useful diagnostic tool for the rapid detection and differentiation of Legionella species in both clinical and epidemiological studies.

AB - Legionella pneumophila serogroup 1 (sg1) accounts for the majority of infections in humans, but other Legionella species are also associated with human disease. In this study, a new SYBR Green I-based multiplex real-time PCR assay in a single reaction was developed to allow the rapid detection and differentiation of Legionella species by targeting specific gene sequences. Candidate target genes were selected, and primer sets were designed by referring to comparative genomic hybridization data of Legionella species. The Legionella species-specific groES primer set successfully detected all 30 Legionella strains tested. The xcpX and rfbA primers specifically detected L. pneumophila sg1-15 and L. pneumophila sg1, respectively. In addition, this assay was validated by testing clinical samples and isolates. In conclusion, this novel multiplex real-time PCR assay might be a useful diagnostic tool for the rapid detection and differentiation of Legionella species in both clinical and epidemiological studies.

KW - Legionella species

KW - Legionnaires' disease

KW - Multiplex

KW - Real-time PCR

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DO - 10.1016/j.mcp.2015.08.004

M3 - Article

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AN - SCOPUS:84951574314

SN - 0890-8508

VL - 29

SP - 414

EP - 419

JO - Molecular and Cellular Probes

JF - Molecular and Cellular Probes

IS - 6

ER -

Multiplex real-time PCR assay for Legionella species

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

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