Recombinase Polymerase Amplification Assay for Simultaneous Detection of Maize Chlorotic Mottle Virus and Sugarcane Mosaic Virus in Maize

Xinran Gao; Yuan Chen; Xuecong Luo; Zhichao Du; Kaiqiang Hao; Mengnan An; Zihao Xia; Yuanhua Wu

doi:10.1021/acsomega.1c01767

Recombinase Polymerase Amplification Assay for Simultaneous Detection of Maize Chlorotic Mottle Virus and Sugarcane Mosaic Virus in Maize

ACS Omega. 2021 Jul 6;6(28):18008-18013. doi: 10.1021/acsomega.1c01767. eCollection 2021 Jul 20.

Authors

Xinran Gao¹, Yuan Chen¹, Xuecong Luo¹, Zhichao Du¹, Kaiqiang Hao¹, Mengnan An¹, Zihao Xia¹, Yuanhua Wu¹

Affiliation

¹ College of Plant Protection, Shenyang Agricultural University, Shenyang, Liaoning 110866, China.

Abstract

Maize chlorotic mottle virus (MCMV) can cause maize lethal necrosis (MLN) when coinfected with potyvirids, such as sugarcane mosaic virus (SCMV), maize dwarf mosaic virus, or wheat streak mosaic virus. MLN is often caused by coinfection of MCMV and SCMV, which has been reported in China and several countries of Africa. In this study, a recombinase polymerase amplification (RPA) assay was established for simultaneous detection of MCMV and SCMV in maize. The RPA assay can be completed within 30 min at 38 °C. The primers for the RPA assay were specific since no crossreaction was detected with other selected viruses that infected maize in China. The detection limit of the RPA method was 10² copies μL^-1, which was about 10-fold more sensitive than that of the conventional PCR method. Moreover, the RPA assay can be successfully applied to detect maize samples collected in the field. These results demonstrated that the established RPA assay is a rapid and efficient method to conduct simultaneous detection of MCMV and SCMV, which provides an alternative technology for MLN diagnosis.