title: Detection and differentiation of Japanese encephalitis virus genotype I and genotype III by reverse transcription loop-mediated isothermal amplification combined with restriction fragment length polymorphism.
abstract: Japanese encephalitis (JE), which is a mosquito-borne arboviral infection, is the leading cause of viral encephalitis in Asian countries. The causative agent of JE is Japanese encephalitis virus (JEV), in which the predominant genotype has changed from genotype III (G III) to genotype I (G I). However, a method for the rapid differentiation between JEV G I and G III remains unavailable. This study aimed to establish a rapid JEV genotyping method using reverse transcription loop-mediated isothermal amplification (RT-LAMP). An Spe I site, which was located in the target sequence (C gene) of JEV G III strains but not in JEV G I strains, was selected as the RT-LAMP target. After testing 64 specimens, results showed that RT-LAMP can detect and differentiate JEV G I and G III specifically. Thus, a novel RT-LAMP system for the rapid detection and differentiation of JEV G I and G III was developed successfully.
authors.affiliation: Swine Research Center, Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Ya'an, 625014, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhang
authors.forename: Sanjie
authors.initials: S
authors.lastname: Cao
authors.forename: Rui
authors.initials: R
authors.lastname: Wu
authors.forename: Shuquan
authors.initials: S
authors.lastname: Zhu
authors.forename: Hanyang
authors.initials: H
authors.lastname: Liu
authors.forename: Lei
authors.initials: L
authors.lastname: Yuan
authors.forename: Shuangyan
authors.initials: S
authors.lastname: Shi
authors.forename: Dan
authors.initials: D
authors.lastname: Zhang
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Huang
authors.forename: Xintian
authors.initials: X
authors.lastname: Wen
authors.forename: Yiping
authors.initials: Y
authors.lastname: Wen
authors.forename: Qigui
authors.initials: Q
authors.lastname: Yan
authors.forename: Yong
authors.initials: Y
authors.lastname: Huang
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Ma
mesh_terms: D000818:Animals; D017931:DNA Primers / Q000235:genetics; D004664:Encephalitis Virus, Japanese / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D004672:Encephalitis, Japanese / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D005838:Genotype; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000821:virology*
chemical_list: D017931:DNA Primers