Development of a reverse transcription recombinase-aided amplification assay for detection of Getah virus

Mincai Nie,Yuancheng Zhou,Yao Huang,Xiangang Sun,Huidan Deng,Ling Zhu,Zhiwen Xu

doi:10.1038/s41598-021-99734-7

Abstract

GETV, an arbo-borne zoonotic virus of the genus Alphavirus, which causes diarrhea and reproduction disorders in swine, lead to serious economic losses to the swine industry in China. At present, the existing methods for GETV detection are time-consuming and low sensitivity, so, a rapid, accurate and sensitive GETV detection method is urgently needed. In this study, a fluorescent reverse transcription recombinase-assisted amplification method (RT-RAA) was successfully established for the rapid detection of GETV. The sensitivity of this method to GETV was 8 copies/reaction and 20 TCID50/reaction. No cross-reaction with other viruses. A total of 118 samples were prepared for GETV detection using fluorescent RT-RAA and SYBR Green I RT-qPCR, the coincidence rate of the two methods was 100%. The results suggest that the RT-RAA method is rapid, sensitive and specific for GETV detection and can be applied in the clinical.

Highlights

Getah virus (GETV), an arbo-borne zoonotic virus of the genus Alphavirus, which causes diarrhea and reproduction disorders in swine, lead to serious economic losses to the swine industry in China
Getah virus (GETV), a zoonotic virus belonging to the genus alphavirus, is a plus-strand RNA virus[1]
The combination of reverse transcription recombinase-assisted amplification method (RT-Recombinase aided amplification (RAA))-F2 and RT-RAA-R3 was used as primer for RT-RAA in subsequent experiments

Summary

Introduction

GETV, an arbo-borne zoonotic virus of the genus Alphavirus, which causes diarrhea and reproduction disorders in swine, lead to serious economic losses to the swine industry in China. A fluorescent reverse transcription recombinase-assisted amplification method (RT-RAA) was successfully established for the rapid detection of GETV. The sensitivity of this method to GETV was 8 copies/reaction and 20 TCID50/reaction. The results suggest that the RT-RAA method is rapid, sensitive and specific for GETV detection and can be applied in the clinical. Since GETV was first isolated in Malaysia in 19552, GETV has been found in Australia, China, Japan, Mongolia, Russia and many other countries[3] It mainly causes fever, diarrhea and reproductive disorders in swines, lead to rash, edema and fever in horses[4,5,6]. Fluorescent RAA has the advantages of high sensitivity, strong specificity, high accuracy and Scientific Reports | (2021) 11:20060

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