期刊论文详细信息
Diseases of Aquatic Organisms
Comparison of a novel in situ polymerase chain reaction (ISPCR) method to other methods for white spot syndrome virus (WSSV) detection in Penaeus vannamei
Xu-feng Jian Ling Lu Yong-gui Chen Siu-Ming Chan Jian-guo He1 
关键词: In situ polymerase chain reaction (ISPCR);    Polymerase chain reaction (PCR);    In situ hybridization (ISH);    WSSV;    Penaeus vannamei;    Latent infection;   
DOI  :  10.3354/dao067171
学科分类:生物科学(综合)
来源: Inter-Research
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【 摘 要 】

ABSTRACT: Penaeus vannamei were experimentally injected with white spot syndrome virus (WSSV) and tested for WSSV at different times post-injection (p.i.) by 1-step polymerase chain reaction (PCR), 2-step PCR, in situ hybridization (ISH) and in situ polymerase chain reaction (ISPCR) in order to compare sensitivity of the methods. With 1-step PCR, 4 of 15 shrimp tested positive for WSSV at 12 h p.i., and all tested positive by 24 h p.i. With 2-step PCR, 13 out of 15 samples tested positive at 2 h p.i. and all were positive by 4 h p.i. Using in situ hybridization, 1 sample tested positive at 18 h p.i. and all were positive by 36 h p.i. With ISPCR, 1 out of 5 samples was positive at 2 h p.i. and all were positive by 8 h p.i. Two-step PCR showed the highest sensitivity, followed by ISPCR, 1-step PCR and ISH. Although ISPCR revealed WSSV in 9 of 10 P. vannamei that tested positive for WSSV using 2-step PCR, none of the shrimp examined showed clinical signs of WSSV infection or detectable WSSV with 1-step PCR. The major infected organs were muscle and the hepatopancreas.

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