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Development of cross-priming amplification coupled with vertical flow visualization for rapid detection of infectious spleen and kidney necrosis virus (ISKNV) in mandarin fish, Siniperca chuatsi

文献类型： 外文期刊

作者： Liu, Wenzhi 1 ; Zhou, Yong 1 ; Fan, Yuding 1 ; Jiang, Nan 1 ; Cain, Kenneth 2 ; Zeng, Lingbing 1 ;

作者机构： 1.Chinese Acad Fishery Sci, Yangtze River Fisheries Res Inst, Wuhan 430223, Hubei, Peoples R China

2.Univ Idaho, Dept Fish & Wildlife Sci, Moscow, ID 83843 USA

3.Univ Idaho, Inst Aquaculture Res, Moscow, ID 83843 USA

关键词： Mandarin fish (Siniperca chuatsi);Infectious spleen and kidney necrosis virus (ISKNV);Cross-priming amplification coupled with vertical flow visualization (CPA-VF);Detection

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）

ISSN： 0166-0934

年卷期： 2018 年 253 卷

页码：

收录情况： SCI

摘要： Infectious spleen and kidney necrosis virus (ISKNV) has been recognized as the causative agent of the most serious disease in cultured mandarin fish, Siniperca chuatsi, in China. Disease outbreaks have resulted in substantial losses to the aquaculture industry. Currently, reliable laboratory detection and identification methods are available for this virus. However, rapid detection methods applicable for on-site diagnosis of this infectious agent are unavailable. To address this need, a nearly instrument-free, cost-effective and simple detection method was developed and optimized and incorporates cross priming amplification coupled with vertical flow visualization for rapid identification of ISKNV (ISKNV-CPA-VF). Results show that cross circulation amplification targeting the conserved region of the major capsid protein (MCP) regiment of the ISKNV genome had a sensitivity 10 times greater than traditional PCR at 64 degrees C within 60 min. The optimized concentration of dNTPs and the concentration for Mg2+ were 1.0 mmol/L and 10 mmol/L, respectively. No cross-reactions with other viruses or bacteria were observed. When combined with the nucleic acid strip detection technology, visual detection of ISKNV amplified products was realized within 3-5 min following amplification. The simplicity and nearly instrument-free method for this ISKNV-CPA-VF assay shows great potential for on-site diagnostics of ISKNV infection in Siniperca chuatsi.

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