Detection of Legionella pneumophila by using fluorescence quantitative PCR

Objective To developa specific,rapid and sensitive fluor escence quantitative PCR assay for detection of Legionella pneumophila.Methods The genomic DNA of Legionella pneumophila and Legionella spp was from Chinese Center for Disease Control and Prevention,and then the local strains of Legionella pneumophila and other control strains were obtained from Zhejiang Provincial Center for Disease Control and Prevention.The primers and TaqMan probe were designed in conservative sequence of Legionella pneumophila mip gene,and then the PCR condition and the reaction system were optimized simultaneously.The specificity,sensitivity,reproducibility of the assay were evaluated.Results This assay had high sensitivity and specificity for detecting L.pneumophila but not to the other Legionella spp.and other strains,the sensitivity of this assay was 10 copies/reaction.The whole process from extraction of genomic DNA of strains to assessment of the results could be done in about 2 hours.It was a simple,convenient assay with good reproducibility.Furthermore,this method can also decrease the probability of contamination.Conclusion The fluor escence quantitative PCR provides a specific,rapid and sensitive method for quantitative detection of L.pneumophila.It is helpful for the rapid detection of environment source of pollution,emergency and clinical samples.