Comparison of reverse transcription loop-mediated isothermal amplification, conventional PCR and real-time PCR assays for Japanese encephalitis virus
Identifieur interne : 002849 ( Main/Exploration ); précédent : 002848; suivant : 002850Comparison of reverse transcription loop-mediated isothermal amplification, conventional PCR and real-time PCR assays for Japanese encephalitis virus
Auteurs : Zhiyong Chen [République populaire de Chine] ; Yuxue Liao [République populaire de Chine] ; Xuemei Ke [République populaire de Chine] ; Jie Zhou [République populaire de Chine] ; Yixiong Chen [République populaire de Chine] ; Lulu Gao [République populaire de Chine] ; Qing Chen [République populaire de Chine] ; Shouyi Yu [République populaire de Chine]Source :
- Molecular Biology Reports [ 0301-4851 ] ; 2011-08-01.
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Abstract
Abstract: 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/μl. 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 63°C. The results suggest that the RT-LAMP assay can be applied as a practical molecular diagnostic tool for JEV infection and surveillance.
Url:
DOI: 10.1007/s11033-010-0525-0
Affiliations:
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