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Development of an RT-PCR detection method for mud crab reovirus

文献类型: 外文期刊

作者: Guo, Zhi-Xun 1 ; Weng, Shao-Ping 1 ; Li, Guang 1 ; Chan, Siu-Ming 3 ; He, Jian-Guo 1 ;

作者机构: 1.Zhongshan Sun Yat Sen Univ, Sch Life Sci, State Key Lab Biocontrol, Guangzhou 510275, Guangdong, Peoples R China

2.Chinese Acad Fishery Sci, S China Sea Fisheries Res Inst, Guangzhou 510300, Guangdong, Peoples R China

3.Univ Hong Kong, Sch Biol Sci, Hong Kong, Hong Kong, Peoples R China

关键词: mud crab reovirus;detection method;Scylla serrata;RT-PCR;tissue specificity

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: 2008 年 151 卷 2 期

页码:

收录情况: SCI

摘要: Mud crab reovirus (MCRV) causes high mortality in the Cultivated mud crab. To control better an outbreak Of this Virus, a rapid, specific and sensitive detection method based on RT-PCR was developed. The MCRV detection method was designed based on the one-step and two-step RT-PCR which resulted in the amplification of predicted products of 433 and 304 bp. The method is specific as no cross-reaction was observed between grass carp hemorrhage virus (GCHV), white spot syndrome virus (WSSV), tiger frog virus (TFV), infectious spleen and kidney necrosis virus (ISKNV), fish nervous necrosis virus (NNV) and the MUSCOVY duck virus (MDRV). One-step PCR amplification Could detect 10(-8) mu g of purified MCRV dsRNA, while two-step PCR amplification could detect 10(-9) mu g MCRV dsRNA. At an early stage of infection, MCRV could be detected in the heart, thoracic ganglion, muscle, intestine, gut, hemolymph, gonad and gill, but not in the Stomach of hepatopancreas. However, in the moribund mud crabs, MCRV could be detected in all tissues examined. (c) 2008 Elsevier B.V. All rights reserved.

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