title: Visual Detection of West Nile Virus Using Reverse Transcription Loop-Mediated Isothermal Amplification Combined with a Vertical Flow Visualization Strip.
abstract: West Nile virus (WNV) causes a severe zoonosis, which can lead to a large number of casualties and considerable economic losses. A rapid and accurate identification method for WNV for use in field laboratories is urgently needed. Here, a method utilizing reverse transcription loop-mediated isothermal amplification combined with a vertical flow visualization strip (RT-LAMP-VF) was developed to detect the envelope (E) gene of WNV. The RT-LAMP-VF assay could detect 10(2) copies/μl of an WNV RNA standard using a 40 min amplification reaction followed by a 2 min incubation of the amplification product on the visualization strip, and no cross-reaction with other closely related members of the Flavivirus genus was observed. The assay was further evaluated using cells and mouse brain tissues infected with a recombinant rabies virus expressing the E protein of WNV. The assay produced sensitivities of 10(1.5) TCID50/ml and 10(1.33) TCID50/ml for detection of the recombinant virus in the cells and brain tissues, respectively. Overall, the RT-LAMP-VF assay developed in this study is rapid, simple and effective, and it is therefore suitable for clinical application in the field.
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences Changchun, China.
authors.forename: Zengguo
authors.initials: Z
authors.lastname: Cao
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical SciencesChangchun, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and ZoonosesYangzhou, China.
authors.forename: Hualei
authors.initials: H
authors.lastname: Wang
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical SciencesChangchun, China; Animal Science and Technology College, Jilin Agricultural UniversityChangchun, China.
authors.forename: Lina
authors.initials: L
authors.lastname: Wang
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical SciencesChangchun, China; College of Veterinary Medicine, Jilin UniversityChangchun, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Li
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical SciencesChangchun, China; Changchun SR Biological Technology Co., Ltd., ChangchunChina.
authors.forename: Hongli
authors.initials: H
authors.lastname: Jin
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical SciencesChangchun, China; Key Laboratory of Emergency Detection for Public Health of Zhejiang Province, Zhejiang Provincial Center for Disease Control and PreventionHangzhou, China.
authors.forename: Changping
authors.initials: C
authors.lastname: Xu
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical SciencesChangchun, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and ZoonosesYangzhou, China.
authors.forename: Na
authors.initials: N
authors.lastname: Feng
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences Changchun, China.
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Wang
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences Changchun, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Li
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences Changchun, China.
authors.forename: Yongkun
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences Changchun, China.
authors.forename: Tiecheng
authors.initials: T
authors.lastname: Wang
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical SciencesChangchun, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and ZoonosesYangzhou, China.
authors.forename: Yuwei
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical SciencesChangchun, China; Key Laboratory of Emergency Detection for Public Health of Zhejiang Province, Zhejiang Provincial Center for Disease Control and PreventionHangzhou, China.
authors.forename: Yiyu
authors.initials: Y
authors.lastname: Lu
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical SciencesChangchun, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and ZoonosesYangzhou, China.
authors.forename: Songtao
authors.initials: S
authors.lastname: Yang
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical SciencesChangchun, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and ZoonosesYangzhou, China.
authors.forename: Xianzhu
authors.initials: X
authors.lastname: Xia
keywords: RT-LAMP-VF; West Nile virus; reverse transcription loop-mediated isothermal amplification; visual detection; visualization strip