細菌学および寄生虫学ジャーナル

概要

A Simple Visual Detection Method of Human Zika Virus Using Reverse Transcription Loop Mediated Isothermal Amplification

Yang Li1,2, Zhenzhou Wan3, Yi Zhou2,4, Xia Jin5, Haifeng Shi1* and Chiyu Zhang 2*

Zika virus (ZIKV) is an emerging mosquito-borne pathogen, and belongs to family Flaviviridae, genus Flavivirus. ZIKV infection was associated with microcephaly and male infertility. The surveillance of ZIKA infection is important for human reproductive health, and the control of the virus spread. Development of a fast and sensitive assay for ZIKV detection is urgently needed for China and other Southeast Asian countries. Here, we developed a one-step, single-tube reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for ZIKV detection. The assay has a limit of detection (LOD) of 88 copies of Zika RNA per 25 μl reaction, and can be completed within 20 min when the template input is more than the LOD. Furthermore, it exhibits high specificity and good reproducibility. Importantly, the result can be visualized by colorimetric change when HNB or calcein is used. To evaluate the viability of the assay, a comparison between the RT-LAMP assay and a more conversional RT-qPCR was performed using 20 cell culture supernatants and three urine samples. The ZIKV RT-LAMP showed perfect agreement in detection with the RT-qPCR. These results demonstrate that the newly developed ZIKV RT-LAMP is simple, rapid and well suited for ZIKV surveillance, especially in a resource-limited regions.