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Rapid Detection of Tembusu Virus by Reverse‐Transcription, Loop‐mediated Isothermal Amplification (RT‐LAMP)
Author(s) -
Tang Y.,
Diao Y.,
Yu C.,
Gao X.,
Chen L.,
Zhang D.
Publication year - 2012
Publication title -
transboundary and emerging diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.392
H-Index - 63
eISSN - 1865-1682
pISSN - 1865-1674
DOI - 10.1111/j.1865-1682.2011.01257.x
Subject(s) - reverse transcription loop mediated isothermal amplification , loop mediated isothermal amplification , sybr green i , detection limit , reverse transcription polymerase chain reaction , microbiology and biotechnology , reverse transcriptase , virology , virus , real time polymerase chain reaction , biology , chemistry , polymerase chain reaction , chromatography , gene , messenger rna , dna , biochemistry
Summary A sensitive reverse‐transcription loop‐mediated isothermal amplification (RT‐LAMP) assay was developed for the rapid detection of Tembusu virus (TMUV) infection. The reaction was performed in one step in a single tube at 64°C for 45 min, with SYBR Green I dye added prior to amplification. The detection limit of the RT‐LAMP assay was approximately 10 copies/μl, and no cross‐reaction with other avian viruses was observed. The assay was evaluated further for the diagnosis of TMUV in field samples and compared with conventional RT‐PCR, demonstrating that results of the RT‐LAMP assay corresponded to those of conventional RT‐PCR. In conclusion, RT‐LAMP with SYBR Green I dye was shown to be a sensitive, simple assay for the rapid diagnosis of TMUV infection in ducks.