Two-Step Scheme for Rapid Identification and Differentiation of Legionella pneumophila and Non-Legionella pneumophila Species

• Published in: Journal of Clinical Microbiology Vol. 48; no. 2; pp. 433 - 439
• Main Authors: Zhan, Xiao-Yong, Li, Lian-Qing, Hu, Chao-Hui, Zhu, Qing-Yi
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.02.2010; American Society for Microbiology (ASM)
• Subjects: Bacteriological Techniques - methods; Bacteriology; Base Sequence; Biological and medical sciences; DNA, Bacterial - genetics; DNA, Ribosomal - genetics; Environmental Microbiology; Fundamental and applied biological sciences. Psychology; Genotype; Humans; Legionella - classification; Legionella - genetics; Legionella - isolation & purification; Legionella pneumophila; Legionnaires' Disease - microbiology; Microbiology; Miscellaneous; Molecular Sequence Data; Polymerase Chain Reaction - methods; Restriction Mapping - methods; RNA, Ribosomal, 16S - genetics; Sensitivity and Specificity; China, People's Rep., Guangdong Prov; Bacteria; Legionella pneumophila; Identification; Legionellaceae
• ISSN: 0095-1137; 1098-660X; 1098-660X
• DOI: 10.1128/JCM.01778-09

A rapid two-step scheme based on PCR amplification and enzymatic digestion analysis of a 226-bp fragment of the 16S rRNA gene was developed to identify the Legionella genus by PCR amplification and to differentiate the Legionella pneumophila and non-Legionella pneumophila species by enzymatic digestion analysis. Among 42 ATCC strains (16 strains of L. pneumophila and 26 strains of non-L. pneumophila) and 200 Legionella isolates from environmental water samples, including pools, rivers, lakes, and cooling towers in Guangdong province, 99.59% of L. pneumophila and non-L. pneumophila strains were correctly identified and differentiated by this scheme. The procedure of this two-step identification and differentiation scheme is simple and takes only about 4 h. These results suggest that this two-step scheme provides a simple and convenient method for the rapid identification and differentiation of L. pneumophila and non-L. pneumophila species.