Detection of prawn white spot bacilliform virus by immunoassay with recombinant antigen.
Prawn white spot bacilliform virus (WSBV) is the major pathogen of prawn disease. To develop a sensitive assay for the early detection of this virus, we generated an antibody against a WSBV-specific protein, P204. The p204 gene was cloned from a WSBV cDNA library and expressed in Escherichia coli. The peptide (P204) encoded by p204 was purified, and its antibody raised in mice. IgG fraction of the anti-P204 serum was purified using a Sepharose column and the Fab fragment was obtained by pepsin digestion. An enzyme-linked immunosorbent assay (ELISA) using this Fab fragment was developed to detect the WSBV in prawn tissues. The experiments showed that Fab of anti-P204 antibody is highly specific for WSBV, and displays a sensitivity of 10(5) viral particles/mg prawn tissues. This is the first report using the Fab fragment prepared from a recombinant antigen to detect prawn viruses by ELISA.
The Third Institute of Oceanography, State Oceanic Administration, 361005, Xiamen, People's Republic of China.
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D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D000956:Antigens, Viral / Q000008:administration & dosage / Q000235:genetics* / Q000276:immunology*; D033364:Decapoda / Q000821:virology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D007114:Immunization; D007140:Immunoglobulin Fab Fragments / Q000097:blood / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D014780:Viruses / Q000276:immunology / Q000302:isolation & purification*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D007140:Immunoglobulin Fab Fragments; D011994:Recombinant Proteins