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Development of a quantitative PCR assay for monitoring Streptococcus agalactiae colonization and tissue tropism in experimentally infected tilapia

文献类型： 外文期刊

作者： Su, Y-L 1 ; Feng, J. 2 ; Li, Y-W 1 ; Bai, J-S 3 ; Li, A-X 1 ;

作者机构： 1.Sun Yat Sen Univ, Sch Life Sci, State Key Lab Biocontrol, Key Lab Aquat Prod Safety,Minist Educ, 135 Xingang West St, Guangzhou 510275, Guangdong, Peoples R China

2.Chinese Acad Fishery Sci, South China Sea Fisheries Res Inst, Key Lab South China Sea Fishery Resources Exploit, Minist Agr, Guangzhou, Guangdong, Peoples R China

3.Guangzhou Airport Extry Exit Inspect & Quarantine, Guangzhou, Guangdong, Peoples R China

关键词： bacterial colonization;Oreochromis niloticus;quantitative real-time PCR;Streptococcus agalactiae

期刊名称：JOURNAL OF FISH DISEASES （ 影响因子：2.767； 五年影响因子：2.689 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： Streptococcus agalactiae has become one of the most important emerging pathogens in the aquaculture industry and has resulted in large economic losses for tilapia farms in China. In this study, three pairs of specific primers were designed and tested for their specificities and sensitivities in quantitative real-time polymerase chain reactions (qPCRs) after optimization of the annealing temperature. The primer pair IGS-s/IGS-a, which targets the 16S-23S rRNA intergenic spacer region, was finally chosen, having a detection limit of 8.6 copies of S. agalactiae DNA in a 20 mu L reaction mixture. Bacterial tissue tropism was demonstrated by qPCR in Oreochromis niloticus 5 days postinjection with a virulent S. agalactiae strain. Bacterial loads were detected at the highest level in brain, followed by moderately high levels in kidney, heart, spleen, intestines, and eye. Significantly lower bacterial loads were observed in muscle, gill and liver. In addition, significantly lower bacterial loads were observed in the brain of convalescent O. niloticus 14 days post-injection with several different S. agalactiae strains. The qPCR for the detection of S. agalactiae developed in this study provides a quantitative tool for investigating bacterial tissue tropism in infected fish, as well as for monitoring bacterial colonization in convalescent fish.

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