1.Visual detection of murray valley encephalitis virus by reverse transcription loop-mediated isothermal amplification.
Rui GONG ; Han Hua WANG ; Hong QIN ; Xiao Ping GUO ; Xue Jun MA
Biomedical and Environmental Sciences 2015;28(3):227-230
A sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for rapid visual detection of Murray valley encephalitis virus (MVEV) infection. The reaction was performed in one step in a single tube at 63 °C for 60 min with the addition of the hydroxynaphthol blue (HNB) dye prior to amplification. The detection limit of the RT-LAMP assay was 100 copies per reaction based on 10-fold dilutions of in vitro transcribed RNA derived from a synthetic MVEV DNA template. No cross-reaction was observed with other encephalitis-associated viruses. The assay was further evaluated using spiked cerebrospinal fluid sample with pseudotype virus containing the NS5 gene of MVEV.
Base Sequence
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DNA Primers
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Encephalitis Virus, Murray Valley
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genetics
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isolation & purification
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Limit of Detection
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Transcription, Genetic