A real-time PCR assay for detection and quantification of Lactococcus garvieae.
OBJECTIVE
To develop a rapid, sensitive, specific tool for the detection and quantification of Lactococcus garvieae in food and environmental samples.


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 C(T) value of 36.75 in qPCR. Quantification of L. garvieae vegetative cells was linear (R(2) = 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.


CONCLUSIONS
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.
Department of Microbiology & Research Institute for Translational System Biomics, Chung-Ang University College of Medicine, Seoul, Korea.
M Y
MY
Jung
Y-H
YH
Chang
W
W
Kim
D004783:Environmental Microbiology; D005516:Food Microbiology / Q000379:methods*; D006801:Humans; D017045:Lactococcus / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity
D012336:RNA, Ribosomal, 16S