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

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

2021Scientific Reports11 citationsDOIOpen Access PDF

Abstract

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 TCID 50 /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.

Topics & Concepts

VirologyReverse transcription polymerase chain reactionBiologyAlphavirusReal-time polymerase chain reactionSYBR Green IReverse transcriptaseVirusMolecular biologyPolymerase chain reactionGeneGeneticsGene expressionViral gastroenteritis research and epidemiologyAnimal Disease Management and EpidemiologyVirus-based gene therapy research
Development of a reverse transcription recombinase-aided amplification assay for detection of Getah virus | Litcius