caa a22 4500 605505519 CHVBK 20210128105158.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s00253-015-6759-y doi (NATIONALLICENCE)springer-10.1007/s00253-015-6759-y Improved PCR assay for the species-specific identification and quantitation of Legionella pneumophila in water [Elektronische Daten] [Min Cho, Tae-Young Ahn, Kiseong Joh, Eui Lee, Dong Park] 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. Springer-Verlag Berlin Heidelberg, 2015 Legionella pneumophila nationallicence LysR transcriptional regulator nationallicence Detection nationallicence Real-time PCR nationallicence Cho Min National Academy of Agricultural Science, Rural Development Administration, 560-500, Jeonju, Republic of Korea aut Ahn Tae-Young Department of Microbiology, Dankook University, 330-714, Cheonan, Republic of Korea aut Joh Kiseong Department of Bioscience and Biotechnology, Hankuk University of Foreign Studies, Wangsan, Yongin, 449-791, Gyeonggi, Republic of Korea aut Lee Eui Department of Oral and Maxillofacial Surgery, Korea University Guro Hospital, 152-703, Seoul, Republic of Korea aut Park Dong National Academy of Agricultural Science, Rural Development Administration, 560-500, Jeonju, Republic of Korea aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/21(2015-11-01), 9227-9236 0175-7598 99:21<9227 2015 99 253 https://doi.org/10.1007/s00253-015-6759-y text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6759-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Cho Min National Academy of Agricultural Science, Rural Development Administration, 560-500, Jeonju, Republic of Korea aut NATIONALLICENCE 700 1- Ahn Tae-Young Department of Microbiology, Dankook University, 330-714, Cheonan, Republic of Korea aut NATIONALLICENCE 700 1- Joh Kiseong Department of Bioscience and Biotechnology, Hankuk University of Foreign Studies, Wangsan, Yongin, 449-791, Gyeonggi, Republic of Korea aut NATIONALLICENCE 700 1- Lee Eui Department of Oral and Maxillofacial Surgery, Korea University Guro Hospital, 152-703, Seoul, Republic of Korea aut NATIONALLICENCE 700 1- Park Dong National Academy of Agricultural Science, Rural Development Administration, 560-500, Jeonju, Republic of Korea aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/21(2015-11-01), 9227-9236 0175-7598 99:21<9227 2015 99 253 SWISSBIB 560478992