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《Southwest China Journal of Agricultural Sciences》 2012-01
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Development of Real-time PCR Assay for Detection of Marteilia refringens in Shellfish

XIE Li-ji,XIE Zhi-xun,PANG Yao-shan,LIU Jia-bo,DENG Xian-wen,XIE Zhi-qin (Guangxi Veterinary Research Institute,Guangxi Nanning 530001,China)  
A pair of primers and a TaqMan probe were designed and synthesized according to the conserved gene sequences of Marteilia refringens,and then reaction parameters were optimized to develop a real-time TaqMan-quantitative PCR assay.After having compared the developed real-time PCR assay with that of routine PCR,the results showed that this real-time PCR assay could detect 40 template copies of plasmid DNA,and its sensitivity was 100 times higher than that of the routine PCR.This real-time PCR results were negative after detecting the samples such as Perkinsus sp.,Haplosporidium sp.,Aeromonas hydrophila,Pseudomonas fluorescens,Vibrio parahaemolyticu,Vibrio alginolyticu,Vibrio fluvialis and Vibrio mimicus.As a result of the sensitivity and specificity of the assay with a relatively rapid and simple procedure,the real-time PCR will be useful as a routine assay for the clinical diagnosis of Marteilia refringens infection.
【Fund】: 国家百千万人才工程人选专项资金项目(945200603);; 广西特聘专家专项经费(2011B020);; 广西科技攻关项目(桂科攻0630001-3M)
【CateGory Index】: S944
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