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Development and evaluation of duplex TaqMan real-time PCR assay for detection and differentiation of wide-type and MGF505-2R gene-deleted African swine fever viruses

文献类型： 外文期刊

作者： Guo, Zhenhua 1 ; Li, Kunpeng 2 ; Qiao, Songlin 1 ; Chen, Xin-Xin 1 ; Deng, Ruiguang 1 ; Zhang, Gaiping 1 ;

作者机构： 1.Henan Acad Agr Sci, Key Lab Anim Immunol, Henan Prov Key Lab Anim Immunol, Minist Agr, Zhengzhou, Peoples R China

2.ZhengZhou ZhongDao Biotechnol Co Ltd, Zhengzhou, Peoples R China

3.Henan Agr Univ, Coll Anim Sci & Vet Med, Zhengzhou 450002, Peoples R China

4.Jiangsu Coinnovat Ctr Prevent & Control Important, Yangzhou, Jiangsu, Peoples R China

关键词： African swine fever virus; Duplex real-time PCR; Differential diagnosis; Gene-deleted strains

期刊名称：BMC VETERINARY RESEARCH （ 影响因子：2.741； 五年影响因子：2.955 ）

ISSN：

年卷期： 2020 年 16 卷 1 期

页码：

收录情况： SCI

摘要： Background African swine fever (ASF) is the most important disease to the pigs and cause serious economic losses to the countries with large-scale swine production. Vaccines are recognized as the most useful tool to prevent and control ASF virus (ASFV) infection. Currently, the MGF505 and MGF360 gene-deleted ASFVs or combined with CD2v deletion were confirmed to be the most promising vaccine candidates. Thus, it is essential to develop a diagnosis method to discriminate wide-type strain from the vaccines used. Results In this study, we established a duplex TaqMan real-time PCR based on the B646L gene and MGF505-2R gene. The sequence alignment showed that the targeted regions of primers and probes are highly conserved in the genotype II ASFVs. The duplex real-time assay can specifically detect B646L and MGF505-2R gene single or simultaneously without cross-reaction with other porcine viruses tested. The limit of detection was 5.8 copies and 3.0 copies for the standard plasmids containing B646L and MGF505-2R genes, respectively. Clinical samples were tested in parallel by duplex real-time PCR and a commercial ASFV detection kit. The detection results of these two assays against B646L gene were well consistent. Conclusion We successfully developed and evaluated a duplex TaqMan real-time PCR method which can effectively distinguish the wide type and MGF505 gene-deleted ASFVs. It would be a useful tool for the clinical diagnosis and control of ASF.

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