Development of a Directly Visualized Recombinase Polymerase Amplification-SYBR Green I Method for the Rapid Detection of African Swine Fever Virus
文献类型： 外文期刊
作者： Zhang, Shuai 1 ; Sun, Aijun 1 ; Wan, Bo 1 ; Du, Yongkun 1 ; Wu, Yanan 1 ; Zhang, Angke 1 ; Jiang, Dawei 1 ; Ji, Pengchao; 1 ;
作者机构： 1.Henan Agr Univ, Coll Vet Med, Zhengzhou, Peoples R China
2.Henan Acad Agr Sci, Henan Prov Key Lab Anim Immunol, Minist Agr, Key Lab Anim Immunol, Zhengzhou, Peoples R China
关键词： African swine fever; African swine fever virus; recombinase polymerase amplification; SYBR green; surveillance
期刊名称：FRONTIERS IN MICROBIOLOGY （ 影响因子：5.64； 五年影响因子：6.32 ）
ISSN： 1664-302X
年卷期： 2020 年 11 卷
页码：
收录情况： SCI
摘要： 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 10(3) copies/mu l of ASFV in 15 min at 35 degrees 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.
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