title: Comparison of a novel in situ polymerase chain reaction (ISPCR) method to other methods for white spot syndrome virus (WSSV) detection in Penaeus vannamei.
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.
authors.affiliation: State Key Laboratory for Biocontrol, School of Life Sciences, Zhongshan University, Guangzhou 510275, PR China.
authors.forename: Xu-Feng
authors.initials: XF
authors.lastname: Jian
authors.forename: Ling
authors.initials: L
authors.lastname: Lu
authors.forename: Yong-gui
authors.initials: YG
authors.lastname: Chen
authors.forename: Siu-Ming
authors.initials: SM
authors.lastname: Chan
authors.forename: Jian-guo
authors.initials: JG
authors.lastname: He
mesh_terms: D000818:Animals; D017931:DNA Primers; D004267:DNA Viruses / Q000302:isolation & purification*; D017403:In Situ Hybridization / Q000379:methods*; D033561:Penaeidae / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013997:Time Factors
chemical_list: D017931:DNA Primers