DAO 67:171-176 (2005)  -  doi:10.3354/dao067171

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 Jian1, Ling Lu1, Yong-gui Chen1, Siu-Ming Chan2, Jian-guo He1,*

1State Key Laboratory for Biocontrol, School of Life Sciences, Zhongshan University, Guangzhou 510275, PR China
2Department of Zoology, The University of Hong Kong, Hong Kong SAR, PR China
*Corresponding author. Email:

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.

KEY WORDS: In situ polymerase chain reaction (ISPCR) · Polymerase chain reaction (PCR) · In situ hybridization (ISH) · WSSV · Penaeus vannamei · Latent infection

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