Comparison of reverse transcription loop-mediated isothermal amplification, conventional PCR and real-time PCR assays for Japanese encephalitis virus.
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.
Department of Epidemiology, School of Public Health and Tropical Medicine, Southern Medical University, 1838 Guangzhou North Road, Guangzhou, 510515, Guangdong Province, China.
Zhiyong
Z
Chen
Yuxue
Y
Liao
Xuemei
X
Ke
Jie
J
Zhou
Yixiong
Y
Chen
LuLu
L
Gao
Qing
Q
Chen
Shouyi
S
Yu
D001681:Biological Assay / Q000379:methods*; D004664:Encephalitis Virus, Japanese / Q000235:genetics* / Q000302:isolation & purification*; D005814:Genes, Viral / Q000235:genetics; D006801:Humans; D057230:Limit of Detection; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D048348:Reverse Transcription / Q000235:genetics*; D012680:Sensitivity and Specificity
D012367:RNA, Viral