Visual detection of West Nile virus using reverse transcription loop-mediated isothermal amplification combined with a vertical flow visualization strip

• Main Authors: Zengguo eCao, Hualei eWang, Lina eWang, Ling eLi, Hongli eJin, Na eFeng, Changping eXu, Jianzhong eWang, Qian eLi, Yongkun eZhao, Tiecheng eWang, Yuwei eGao, Yiyu eLu, Songtao eYang, Xianzhu eXia
• Format: Article
• Language: English
• Published: Frontiers Media S.A. 2016-04-01
• Series: Frontiers in Microbiology
• Subjects: West Nile virus; visual detection; Reverse transcription loop-mediated isothermal amplification; visualization strip; RT-LAMP-VF
• Online Access: http://journal.frontiersin.org/Journal/10.3389/fmicb.2016.00554/full

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 methodfor 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 102 copies/μl ofan WNV RNA standard using a 40 min amplification reaction followed by a 2 min incubationof the amplification product on the visualization strip, and no cross-reaction with other closely related members of theFlavivirus 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 101.5TCID50/ml and 101.33 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.