title: Improvement of immunodetection of white spot syndrome virus using a monoclonal antibody specific for heterologously expressed icp11.
abstract: The icp11 gene encoding the highly abundant DNA mimic protein of white spot syndrome virus (WSSV) was cloned into the pTYB1 and pGEX-6P-1 expression vectors and introduced into E. coli by transformation. After induction, C-terminally intein-tagged ICP11 (ICP11-intein) and N-terminally glutathione-S-transferase (GST)-tagged ICP11 (GST-ICP11) proteins with molecular masses of 64 and 35 kDa were obtained. These proteins were purified by SDS-PAGE and used for immunization of Swiss mice for monoclonal antibody (MAb) production. Two MAbs specific for ICP11 were selected; these MAbs can be used to detect natural WSSV infection in Penaeus vannamei by dot blotting, western blotting or immunohistochemistry without cross-reaction with other shrimp tissues or other common shrimp viruses. The detection sensitivity of the MAbs was approximately 0.7 fmole/spot of GST-ICP11 as determined by dot blotting. These MAbs showed stronger immunoreactivity than other MAbs from previous studies that are specific for VP28 and VP19. A combination of MAbs specific for ICP11, VP28 and VP19 increased the detection sensitivity of WSSV during early infection to a sensitivity 250 times lower than that of one-step PCR. Therefore, the MAbs specific for ICP11 could be used to confirm and enhance the detection sensitivity for WSSV infection in shrimp using various types of antibody-based assays.
authors.affiliation: Department of Biology, Srinakharinwirot University, Sukhumvit 23, Bangkok 10110, Thailand.
authors.forename: Ruthairat
authors.initials: R
authors.lastname: Siriwattanarat
authors.forename: Siwaporn
authors.initials: S
authors.lastname: Longyant
authors.forename: Parin
authors.initials: P
authors.lastname: Chaivisuthangkura
authors.forename: Pradit
authors.initials: P
authors.lastname: Wangman
authors.forename: Akapon
authors.initials: A
authors.lastname: Vaniksampanna
authors.forename: Paisarn
authors.initials: P
authors.lastname: Sithigorngul
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000302:isolation & purification; D000956:Antigens, Viral / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D015151:Immunoblotting / Q000379:methods; D007150:Immunohistochemistry / Q000379:methods; D051379:Mice; D033561:Penaeidae / Q000821:virology; D012680:Sensitivity and Specificity; D014730:Veterinary Medicine / Q000379:methods*; D014773:Virology / Q000379:methods*; D046848:White spot syndrome virus 1 / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral