Improved PCR assay for the species-specific identification and quantitation of Legionella pneumophila in water

Verfasser / Beitragende:
[Min Cho, Tae-Young Ahn, Kiseong Joh, Eui Lee, Dong Park]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/21(2015-11-01), 9227-9236
Format:
Artikel (online)
ID: 605505519
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024 7 0 |a 10.1007/s00253-015-6759-y  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-015-6759-y 
245 0 0 |a Improved PCR assay for the species-specific identification and quantitation of Legionella pneumophila in water  |h [Elektronische Daten]  |c [Min Cho, Tae-Young Ahn, Kiseong Joh, Eui Lee, Dong Park] 
520 3 |a Legionellosis outbreak is a major global health care problem. However, current Legionella risk assessments may be compromised by uncertainties in Legionella detection methods, infectious dose, and strain infectivity. These limitations may place public health at significant risk, leading to significant monetary losses in health care. However, there are still unmet needs for its rapid identification and monitoring of legionellae in water systems. Therefore, in the present study, a primer set was designed based on a LysR-type transcriptional regulator (LTTR) family protein gene of Legionella pneumophila subsp. pneumophila str. Philadelphia 1 because it was found that this gene is structurally diverse among species through BLAST searches. The specificity of the primer set was evaluated using genomic DNA from 6 strains of L. pneumophila, 5 type strains of other related Legionella species, and other 29 reference pathogenic bacteria. The primer set used in the PCR assay amplified a 264-bp product for only targeted six strains of L. pneumophila. The assay was also able to detect at least 1.39 × 103 copies/μl of cloned amplified target DNA using purified DNA or 7.4 × 100 colony-forming unit per reaction when using calibrated cell suspension. In addition, the sensitivity and specificity of this assay were confirmed by successful detection of Legionella pneumophila in environmental water samples. 
540 |a Springer-Verlag Berlin Heidelberg, 2015 
690 7 |a Legionella pneumophila  |2 nationallicence 
690 7 |a LysR transcriptional regulator  |2 nationallicence 
690 7 |a Detection  |2 nationallicence 
690 7 |a Real-time PCR  |2 nationallicence 
700 1 |a Cho  |D Min  |u National Academy of Agricultural Science, Rural Development Administration, 560-500, Jeonju, Republic of Korea  |4 aut 
700 1 |a Ahn  |D Tae-Young  |u Department of Microbiology, Dankook University, 330-714, Cheonan, Republic of Korea  |4 aut 
700 1 |a Joh  |D Kiseong  |u Department of Bioscience and Biotechnology, Hankuk University of Foreign Studies, Wangsan, Yongin, 449-791, Gyeonggi, Republic of Korea  |4 aut 
700 1 |a Lee  |D Eui  |u Department of Oral and Maxillofacial Surgery, Korea University Guro Hospital, 152-703, Seoul, Republic of Korea  |4 aut 
700 1 |a Park  |D Dong  |u National Academy of Agricultural Science, Rural Development Administration, 560-500, Jeonju, Republic of Korea  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/21(2015-11-01), 9227-9236  |x 0175-7598  |q 99:21<9227  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-015-6759-y  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-015-6759-y  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Cho  |D Min  |u National Academy of Agricultural Science, Rural Development Administration, 560-500, Jeonju, Republic of Korea  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Ahn  |D Tae-Young  |u Department of Microbiology, Dankook University, 330-714, Cheonan, Republic of Korea  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Joh  |D Kiseong  |u Department of Bioscience and Biotechnology, Hankuk University of Foreign Studies, Wangsan, Yongin, 449-791, Gyeonggi, Republic of Korea  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Lee  |D Eui  |u Department of Oral and Maxillofacial Surgery, Korea University Guro Hospital, 152-703, Seoul, Republic of Korea  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Park  |D Dong  |u National Academy of Agricultural Science, Rural Development Administration, 560-500, Jeonju, Republic of Korea  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/21(2015-11-01), 9227-9236  |x 0175-7598  |q 99:21<9227  |1 2015  |2 99  |o 253 
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