Multiplex real-time PCR assay for Legionella species.
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.
Department of Medicine, College of Medicine, Korea University, Seoul, Republic of Korea.
Seung Min
SM
Kim
Institute of Emerging Infectious Diseases, Korea University College of Medicine, Seoul, Republic of Korea.
Yoojung
Y
Jeong
Division of Infectious Diseases, Department of Internal Medicine, Korea University College of Medicine, Seoul, Republic of Korea; Institute of Emerging Infectious Diseases, Korea University College of Medicine, Seoul, Republic of Korea.
Jang Wook
JW
Sohn
Department of Medicine, College of Medicine, Korea University, Seoul, Republic of Korea; Division of Infectious Diseases, Department of Internal Medicine, Korea University College of Medicine, Seoul, Republic of Korea; Institute of Emerging Infectious Diseases, Korea University College of Medicine, Seoul, Republic of Korea. Electronic address: macropha@korea.ac.kr.
Min Ja
MJ
Kim
D052160:Benzothiazoles; D055028:Comparative Genomic Hybridization; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis; D003959:Diamines; D006801:Humans; D007875:Legionella / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D007877:Legionnaires' Disease / Q000175:diagnosis*; D025202:Molecular Diagnostic Techniques / Q000379:methods; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009930:Organic Chemicals / Q000378:metabolism; D011804:Quinolines; D012680:Sensitivity and Specificity; D013183:Sputum / Q000382:microbiology
D052160:Benzothiazoles; D017931:DNA Primers; D004269:DNA, Bacterial; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I
Legionella species; Legionnaires' disease; Multiplex; Real-time PCR