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RT ‐ LAMP detection of shrimp Taura syndrome virus ( TSV ) by combination with a nanogold‐oligo probe
Author(s) -
Phromjai Jurairat,
Mathuros Thitima,
Phokharatkul Ditsayuth,
Prombun Potchanathorn,
Suebsing Rungkarn,
Tuantrat Adisorn,
Kiatpathomchai Wansika
Publication year - 2015
Publication title -
aquaculture research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.646
H-Index - 89
eISSN - 1365-2109
pISSN - 1355-557X
DOI - 10.1111/are.12345
Subject(s) - loop mediated isothermal amplification , amplicon , biology , shrimp , microbiology and biotechnology , reverse transcription loop mediated isothermal amplification , detection limit , colloidal gold , dna , virology , chromatography , polymerase chain reaction , reverse transcriptase , gene , materials science , nanotechnology , nanoparticle , biochemistry , chemistry , fishery
This study describes the application and optimization of a newly developed specific gold nanoparticle (Au NP ) visual detection method to detect a reverse transcription loop‐mediated isothermal amplification ( RT ‐ LAMP ) product of Taura syndrome virus ( TSV ) in shrimp with simple and cheaper in comparison to conventional gel electrophoresis. Briefly, the visual detection method is based on prevention of a salt induced DNA ‐labelled Au NP colour change from red to blue–purple due to hybridization with complementary DNA amplicons that arise from a positive TSV RT ‐ LAMP reaction. RT ‐ LAMP combined with visual amplicon detection using the DNA ‐labelled Au NP ‐probe had high sensitivity and the probe did not cross react with amplicons obtained using specific LAMP methods with other shrimp viruses. The advantages of this assay include simplicity, short analysis time and low‐cost, suitable for field laboratory applications.