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Semiquantitative dot‐blot immunogold assay for specific detection of white spot syndrome virus
Author(s) -
Bunsag Nittaya,
Chotigeat Wilaiwan,
Deachamag Panchalika,
Thananimit Suchera
Publication year - 2018
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1640
Subject(s) - white spot syndrome , dot blot , colloidal gold , nitrocellulose , immunogold labelling , microbiology and biotechnology , virus , polyclonal antibodies , biology , chemistry , virology , antibody , chromatography , nanoparticle , membrane , dna , materials science , biochemistry , nanotechnology , immunology
A dot‐blot immunogold assay (DBIA) was developed to detect white spot syndrome virus (WSSV) using the polyclonal antibody VP26 (anti‐VP26). The anti‐VP26 was immobilized on gold nanoparticles (Ab‐AuNPs), and a nitrocellulose membrane was used as a detection pad. When the target WSSV bound to the Ab‐AuNPs a reddish dot appeared on the surface of the membrane used within 2–5 Min, which could be seen with the naked eye. The test was able to detect WSSV at concentrations as low as 10 5 copies μL −1 of WSSV. The DBIA developed had good specificity, and the colloidal gold probe can be applied within 2–3 days when stored at 4 °C. For real sample analysis, the DBIA was applied to samples of seawater used for shrimp cultivation without sample preparation. The results indicate that sample 1 showed a positive result, whereas samples 2 and 3 produced negative results. Then, samples 2 and 3 were spiked with WSSV for method validation. To confirm the performance of the DBIA developed, polymerase chain reaction (PCR) was conducted and the PCR results were the same as those found by the DBIA. Therefore, the DBIA developed could be applied for WSSV detection in real water samples.