Validation of a new seminested PCR-based detection method for Legionella pneumophila

• Published in: Journal of microbiological methods Vol. 70; no. 1; pp. 214 - 217
• Main Authors: YANEZ, M. A, BARBERA, V. M, CATALAN, V
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier B.V 01-07-2007; Elsevier Science
• Subjects: Bacterial Proteins - genetics; Bacteriological methods and techniques used in bacteriology; Bacteriological Techniques; Bacteriology; Biological and medical sciences; Carrier Proteins - genetics; Environmental samples; Fundamental and applied biological sciences. Psychology; Legionella pneumophila; Legionella pneumophila - genetics; Legionella pneumophila - isolation & purification; Membrane Proteins - genetics; Method validation; Microbiology; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - standards; Reference Standards; Seminesled PCR; Sensitivity and Specificity; Water Microbiology; Legionella pneumophila; Environmental samples; Method validation; Seminesled PCR; Legionellaceae; Sample; Legionellosis; Method; Infection; Polymerase chain reaction; Bacteriosis; Bacteria; Environment; Detection
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/j.mimet.2007.03.013

We report a new seminested PCR method for detection of
Legionella pneumophila based on the simultaneous amplification of a 387 bp fragment of the
dotA gene and a 827 bp recombinant fragment that serves as an internal positive control. This new detection system was validated and its specificity and detection limit were determined. Parallel analysis of 90 environmental samples to compare this method, a real-time PCR method and the standard culture isolation method, demonstrated that this seminested method is useful for the study of
L. pneumophila.