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Development of real-time PCR for detection and quantitation of Streptococcus parauberis

Nguyen TL, Lim Y, Kim D & Austin B (2016) Development of real-time PCR for detection and quantitation of Streptococcus parauberis, Journal of Fish Diseases, 39 (1), pp. 31-39.

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.

gyrB; qPCR; Real-time PCR; Streptococcus parauberis; TaqMan probe

AuthorsNguyen Thanh Luan, Lim Yun-Jin, Kim Do-Hyung, Austin Brian
Publication date01/2016
Publication date online27/10/2014
Date accepted by journal07/09/2014
ISSN 0140-7775

Journal of Fish Diseases: Volume 39, Issue 1 (2016)

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