A real-time PCR assay for detection and quantification of Lactococcus garvieae
Article first published online: 29 SEP 2009
© 2009 The Authors. Journal compilation © 2009 The Society for Applied Microbiology
Journal of Applied Microbiology
Volume 108, Issue 5, pages 1694–1701, May 2010
How to Cite
Jung, M.Y., Chang, Y.-H. and Kim, W. (2010), A real-time PCR assay for detection and quantification of Lactococcus garvieae. Journal of Applied Microbiology, 108: 1694–1701. doi: 10.1111/j.1365-2672.2009.04568.x
- Issue published online: 8 APR 2010
- Article first published online: 29 SEP 2009
- 2009/0965: received 2 June 2009, revised 18 August 2009 and accepted 15 September 2009
- 16S rRNA;
- quantitative polymerase chain reaction
Aims: To develop a rapid, sensitive, specific tool for the detection and quantification of Lactococcus garvieae in food and environmental samples.
Methods and Results: A real-time quantitative PCR (qPCR) assay with primers for CAU12F and CAU12R based on the 16S rRNA gene of L. garvieae was successfully established. The limit of detection for L. garvieae genomic DNA was 1 ng DNA in conventional PCR and 32 fg with a mean CT value of 36·75 in qPCR. Quantification of L. garvieae vegetative cells was linear (R2 = 0·99) over a 7-log-unit dynamic range down to ten L. garvieae cells.
Conclusions: This method is highly specific, sensitive and reproducible for the detection of L. garvieae compared to gel-based conventional PCR assays, thus providing precise quantification of L. garvieae in food and natural environments.
Significance and Impact of the Study: This work provides efficient diagnostic and monitoring tools for the rapid identification of L. garvieae, an emerging pathogen in aquaculture and an occasional human pathogen from other members of the genus Lactobacillus.