NDLI: Development of a Directly Visualized Recombinase Polymerase Amplification–SYBR Green I Method for the Rapid Detection of African Swine Fever Virus

Content Provider	Directory of Open Access Journals (DOAJ)
Author	Shuai Zhang Aijun Sun Bo Wan Yongkun Du Yanan Wu Angke Zhang Dawei Jiang Pengchao Ji Zhanyong Wei Guoqing Zhuang Gaiping Zhang
Abstract	African swine fever (ASF) is a lethal disease in swine caused by etiologic African swine fever virus (ASFV). The global spread of ASFV has resulted in huge economic losses globally. In the absence of effective vaccines or drugs, pathogen surveillance has been the most important first-line intervention to prevent ASF outbreaks. Among numerous diagnostic methods, recombinase polymerase amplification (RPA)-based detection is capable of producing sensitive and specific results without relying on the use of expensive instruments. However, currently used gene-specific, probe-based RPA for ASFV detection is expensive and time-consuming. To improve the efficiency of ASFV surveillance, a novel directly visualized SYBR Green I-staining RPA (RPAS) method was developed to detect the ASFV genome. SYBR Green I was added to the amplified RPA products for direct visualization by the naked eye. The sensitivity and specificity of this method were confirmed using standard plasmid and inactivated field samples. This method was shown to be highly specific with a detection limit of 103 copies/μl of ASFV in 15 min at 35°C without any cross-reactions with other important porcine viruses selected. In summary, this method enables direct sample visualization with reproducible results for ASFV detection and hence has the potential to be used as a robust tool for ASF prevention and control.
e-ISSN	1664302X
DOI	10.3389/fmicb.2020.602709
Journal	Frontiers in Microbiology
Volume Number	11
Language	English
Publisher	Frontiers Media S.A.
Publisher Date	2020-12-01
Publisher Place	Switzerland
Access Restriction	Open
Subject Keyword	Microbiology African Swine Fever African Swine Fever Virus Recombinase Polymerase Amplification Sybr Green Surveillance
Content Type	Text
Resource Type	Article

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