RRC ID 48453
Author Nguyen TL, Lim YJ, Kim DH, Austin B.
Title Development of real-time PCR for detection and quantitation of Streptococcus parauberis.
Journal J Fish Dis
Abstract Streptococcus parauberis is an increasing threat to aquaculture of olive flounder, Paralichthys olivaceus Temminck & Schlegel, in South Korea. We developed a real-time polymerase chain reaction (PCR) method using the TaqMan probe assay to detect and quantify S. parauberis by targeting the gyrB gene sequences, which are effective for molecular analysis of the genus Streptococcus. Our real-time PCR assay is capable of detecting 10 fg of genomic DNA per reaction. The intra- and interassay coefficient of variation (CV) values ranged from 0.42-1.95%, demonstrating that the assay has good reproducibility. There was not any cross-reactivity to Streptococcus iniae or to other streptococcal/lactococcal fish pathogens, such as S. agalactiae and Lactococcus garvieae, indicating that the assay is highly specific to S. parauberis. The results of the real-time PCR assay corresponded well to those of conventional culture assays for S. parauberis from inoculated tissue homogenates (r = 0.957; P < 0.05). Hence, this sensitive and specific real-time PCR is a valuable tool for diagnostic quantitation of S. parauberis in clinical samples.
Volume 39(1)
Pages 31-9
Published 2016-1-1
DOI 10.1111/jfd.12322
PMID 25345976
MeSH Animals DNA Gyrase / genetics DNA Primers / standards DNA, Bacterial / analysis Fish Diseases / microbiology* Fisheries Flounder* Fluorescent Dyes / standards Gene Dosage Intestinal Mucosa / microbiology Kidney / microbiology Real-Time Polymerase Chain Reaction / standards Real-Time Polymerase Chain Reaction / veterinary* Reproducibility of Results Sensitivity and Specificity Spleen / microbiology Streptococcal Infections / microbiology Streptococcal Infections / veterinary* Streptococcus / genetics Streptococcus / growth & development Streptococcus / isolation & purification*
IF 2.318
Times Cited 10
General Microbes JCM 5709