Journal
MOLECULAR BIOLOGY REPORTS
Volume 38, Issue 6, Pages 4063-4070Publisher
SPRINGER
DOI: 10.1007/s11033-010-0525-0
Keywords
Japanese encephalitis virus; Reverse transcription loop-mediated isothermal amplification (RT-LAMP); Real-time RT-PCR; RT-PCR
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Funding
- National Natural Science Foundation of China [30972525]
- Guangdong Province 211 Project [GW201004]
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We developed and evaluated a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detecting Japanese encephalitis virus (JEV). The sensitivity of the JEV RT-LAMP assay was in concordance with that of real-time RT-PCR and 10-fold more sensitive than that of conventional RT-PCR, which the detection limit was 24 copies/mu l. The JEV RT-LAMP was highly specific, which no cross-reactivity was found with dengue-2 virus, rabies virus, norovirus, astrovirus and human enterovirus 71. The JEV RT-LAMP assay was more simple and less time-consuming compared to the conventional RT-PCR and real-time RT-PCR, which the amplification could be completed in a single tube within 1 h under isothermal conditions at temperature of 63A degrees C. The results suggest that the RT-LAMP assay can be applied as a practical molecular diagnostic tool for JEV infection and surveillance.
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