Temporal analysis of VP28 gene of white spot syndrome virus (WSSV) was carried out using RT-PCR and Western blot in different hosts (marine shrimp, Penaeus indicus and Metapenaeus monoceros, freshwater prawn, Macrobrachium rosenbergii and freshwater crab, Paratelphusa hydrodomous) with different habitats and response to WSSV for comparison. The VP28 gene from Indian WSSV isolate was sequenced and the sequence was deposited in GenBank (accession no. AY422228). RT-PCR analysis was used to detect the VP28-specific transcript in DNase treated total RNA from hemolymph and gill tissue from WSSV-infected P. indicus, M. monoceros, M. rosenbergii and P. hydrodomous before infection (0 h) and at different time intervals of post-infection. In P. indicus and M. monoceros, the WSSV caused 100% mortality at 72 h p.i. and transcript was detected at 12 h p.i. whereas in freshwater crab the transcript was detected at 2 days p.i. with 100% mortality at 84 h p.i. The expression of this protein was also detected first in P. indicus and M. monoceros at 24 h p.i., and in P. hydrodomous at 72 h p.i. These observations correlate with the results of mortality data. © 2005 Elsevier B.V. All rights reserved.