title: Detection of white spot syndrome virus (WSSV) of shrimp by means of monoclonal antibodies (MAbs) specific to an envelope protein (28 kDa).
abstract: The vp28 gene encoding an envelope protein (28 kDa) of white spot syndrome virus (WSSV) was amplified from WSSV-infected tiger shrimp that originated from Malaysia. Recombinant VP28 protein (r-28) was expressed in Escherichia coli and used as an antigen for preparation of monoclonal antibodies (MAbs). Three murine MAbs (6F6, 6H4 and 9C10) that were screened by r-28 antigen-based enzyme-linked immunosorbent assay (ELISA) were also able to recognize viral VP28 protein as well as r-28 on Western blot. Three non-overlapping epitopes of VP28 protein were determined using the MAbs in competitive ELISA; thus, an antigen-capture ELISA (Ac-ELISA) was developed by virtue of these MAbs. Ac-ELISA can differentiate WSSV-infected shrimp from uninfected shrimp and was further confirmed by a polymerase chain reaction (PCR) and Western blot. Approximately 400 pg of purified WSSV sample and 20 pg of r-28 could be detected by Ac-ELISA, which is comparable in sensitivity to PCR assay but more sensitive than Western blot in the detection of purified virus. Hemolymph and tissue homogenate samples collected from a shrimp farm in Malaysia during December 2000 and July 2001 were also detected by Ac-ELISA and PCR with corroborating results.
authors.affiliation: Institute of Molecular Agrobiology, The National University of Singapore, Singapore.
authors.forename: W
authors.initials: W
authors.lastname: Liu
authors.forename: Y T
authors.initials: YT
authors.lastname: Wang
authors.forename: D S
authors.initials: DS
authors.lastname: Tian
authors.forename: Z C
authors.initials: ZC
authors.lastname: Yin
authors.forename: J
authors.initials: J
authors.lastname: Kwang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000956:Antigens, Viral / Q000032:analysis*; D017756:Aquaculture; D015153:Blotting, Western / Q000379:methods / Q000662:veterinary; D004267:DNA Viruses / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D004279:DNA, Viral / Q000032:analysis; D033364:Decapoda / Q000821:virology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary; D005784:Gene Amplification; D008296:Malaysia; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D012680:Sensitivity and Specificity; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D004279:DNA, Viral; D014759:Viral Envelope Proteins