Development of immunodot blot assay for the detection of white spot syndrome virus infection in shrimps (Penaeus monodon)
作者： Mudagandur S Shekhar , Gopalapillay Gopikrishna
DOI: 10.1111/J.1365-2109.2010.02553.X
关键词:
摘要: The VP 28 gene encoding a structural envelope protein of the white spot syndrome virus (WSSV) was cloned into pET32a(+) expression vector for production recombinant VP28 protein. A purified 39.9 kDa size used polyclonal antibody in rabbit. Specific immunoreactivity rabbit anti rVP28 antiserum to viral antigen confirmed by Western blot. specificity this anti-rVP28 detect presence WSSV-infected Penaeus monodon verified using immunodot blot assay. Immunodot showed positive reaction infected shrimp tissues with prominent colour development 3,3′,5,5′-tetramethylbenzidine (TMB) as chromogenic substrate when compared 3–3′ diaminobenzidine tetrahydrochloride (DAB). Highest signal intensities immunodots were observed pleopod extracts and haemolymph. On comparison polymerase chain (PCR), could 76% PCR-positive samples. found be equivalent first-step PCR sensitivity WSSV particles estimated contain 1.0 × 105 viral DNA copies.
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