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Development of a nitrocellulose‐enzyme immunoassay for the detection of yellow‐head virus from penaeid shrimp
Author(s) -
Lu Y.,
Tapay L. M.,
Loh P. C.
Publication year - 1996
Publication title -
journal of fish diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 85
eISSN - 1365-2761
pISSN - 0140-7775
DOI - 10.1111/j.1365-2761.1996.tb00114.x
Subject(s) - horseradish peroxidase , nitrocellulose , immunoassay , shrimp , biology , virus , peroxidase , enzyme , virology , microbiology and biotechnology , antibody , biochemistry , fishery , immunology , membrane
A nitrocellulose‐enzyme immunoassay (NC‐EIA) employing horseradish peroxidase (HRP) was developed for the detection of yellow‐head virus in the tissues of infected penaeid shrimp. Two substrates, DAB (3,3¢‐diaminobenzidine retrahydrochloride) and TMB (3,3¢5,5¢‐tetramethylbenzidine) for HRP were evaluated. The TMB was found to produce a more distinct and easily recognizable colour product and provided enhanced sensitivity (0.4 ng for TMB versus 0.8 ng viral protein for DAB). Under the present format, the NC‐EIA was capable of detecting as little as 100 TCID 50 of virus in infected tissues. Uninfected tissue homogenates yielded no significant background colouration.