title: Rapid and simple detection of Japanese encephalitis virus by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick.
abstract: Japanese encephalitis virus (JEV) is a major cause of viral encephalitis in geographical areas, such as Asia and Western Pacific, where it is a threat to human and animal health. To control this disease, it is necessary to develop a rapid, simple, accurate method for diagnosis. In this study, a method based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) coupled with a lateral flow dipstick (LFD) has been developed to detect JEV (JEV RT-LAMP-LFD). The entire assay can be completed within 70 min, and in this study, no false positive results were observed when other pathogens were tested, indicating that the assay is a highly specific method for the detection of JEV. Additionally, the sensitivity of the RT-LAMP-LFD assay for SA14-14-2 strain was 50 pg of RNA, which was similar to that of RT-PCR and RT-LAMP combined with gel electrophoresis, and was 10-fold more sensitive than RT-LAMP combined with calcein. The limit of detection for this assay was 5 pg of RNA. In addition, no false positive results were obtained with 14 serum samples. Our results indicate that this RT-LAMP-LFD assay will be of great value for JEV infection testing due to its rapid and highly specific and sensitive properties.
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
authors.forename: Jieru
authors.initials: J
authors.lastname: Deng
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Pei
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
authors.forename: Hongchao
authors.initials: H
authors.lastname: Gou
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
authors.forename: Zuodong
authors.initials: Z
authors.lastname: Ye
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
authors.forename: Cuicui
authors.initials: C
authors.lastname: Liu
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, China. Electronic address: jdchen@scau.edu.cn.
authors.forename: Jinding
authors.initials: J
authors.lastname: Chen
mesh_terms: D000818:Animals; D001208:Asia; D002846:Chromatography, Affinity / Q000379:methods*; D004664:Encephalitis Virus, Japanese / Q000235:genetics / Q000302:isolation & purification*; D004672:Encephalitis, Japanese / Q000175:diagnosis / Q000662:veterinary; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D013696:Temperature; D013997:Time Factors
keywords: Japanese encephalitis virus (JEV); Lateral flow dipstick (LFD); Reverse transcription loop-mediated isothermal amplification (RT-LAMP)