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LAMP-based method for a rapid identification of Legionella spp. and Legionella pneumophila

  • Xi Lu
  • Zi-Yao Mo
  • Hong-Bo Zhao
  • He Yan
  • Lei Shi
Methods and Protocols

Abstract

Legionella pneumophila is accounted for more than 80% of Legionella infection. However it is difficult to discriminate between the L. pneumophila and non-L. pneumophila species rapidly. In order to detect the Legionella spp. and distinguish L. pneumophila from Legionella spp., a real-time loop-mediated isothermal amplification (LAMP) platform that targets a specific sequence of the 16S rRNA gene was developed. LS-LAMP amplifies the fragment of the 16S rRNA gene to detect all species of Legionella genus. A specific sequence appears at the 16S rRNA gene of L. pneumophila, while non-L. pneumophila strains have a variable sequence in this site, which can be recognized by the primer of LP-LAMP. In the present study, 61 reference strains were used for the method verification. We found that the specificity was 100% for both LS-LAMP and LP-LAMP, and the sensitivity of LAMP assay for L. pneumophila detection was between 52 and 5.2 copies per reaction. In the environmental water samples detection, a total of 107 water samples were identified by the method. The culture and serological test were used as reference methods. The specificity of LS-LAMP and LP-LAMP for the samples detection were 91.59% (98/107) and 93.33% (56/60), respectively. The sensitivity of LS-LAMP and LP-LAMP were 100% (51/51) and 100% (18/18). The results suggest that real-time LAMP, as a new assay, provides a specific and sensitive method for rapid detection and differentiation of Legionella spp. and L. pneumophila and should be utilized to test environmental water samples for increased rates of detection.

Keywords

Loop-mediated isothermal amplification Legionella pneumophila Different diagnosis 

Notes

Acknowledgments

The project was funded by the Science and Technology Development Fund of Macao (039/2007/A3), the National Natural Science Foundation of China (20877028), and the State Key Laboratory of Respiratory Diseases (2007DA780154F0904).

We also thank Qing-Yi Zhu from the Guangzhou Kingmed Center for Clinical Laboratory for providing the DNA or strains of L. pneumophila and non-L. pneumophila.

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Xi Lu
    • 1
  • Zi-Yao Mo
    • 2
  • Hong-Bo Zhao
    • 2
  • He Yan
    • 1
  • Lei Shi
    • 1
  1. 1.College of Light Industry and Food SciencesSouth China University of TechnologyGuangzhouChina
  2. 2.The State Key Laboratory of Respiratory DiseasesGuangzhou Medical UniversityGuangzhouChina

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