Development and evaluation of multi-epitope protein p72 (MeP72) for the serodiagnosis of African swine fever.

IF 1.1 4区医学 Q4 VIROLOGY

Acta virologica Pub Date : 2021-01-01 DOI:10.4149/av_2021_304

Xingxing Zhang, Jing Guo, Lixia Wang, Zhiyuan Li, Yucheng Liu, Lulu Tian, Chencheng Xiao, Yanfang Li, Xuepeng Cai, Qingling Meng, Jun Qiao

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引用次数: 2

Abstract

African swine fever (ASF) is an acute and severe infectious disease that seriously endangers the global porcine industry. In order to develop ASF serodiagnostic reagents with high specificity and sensitivity, in the present study, the antigenic epitopes of P72 protein of African swine fever virus (ASFV) were analyzed, and the ASFV multi-epitope fusion gene MeP72 in tandem with the dominant linear epitopes was constructed. The recombinant multi-epitope fusion MeP72 (reMeP72) was prepared in Escherichia coli. A colloidal gold-based immunochromatographic assay (CGIA) based on reMeP72 was developed for the detection of antibodies against ASFV. A total of 139 pig clinical serum samples were used for assessment of the potential diagnostic value of reMeP72. The results showed that CGIA did not cross-react with positive sera of viruses, such as classical swine fever virus (CSFV), porcine circovirus type 2 (PCV2), pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV) and swine influenza virus (SIV), showing high specificity. Sensitivity analysis showed that CGIA could detect ASFV-positive serum at a dilution of 1:64. Compared with commercial ASFV kits, the sensitivity and specificity of ASFV CGIA based on reMeP72 protein were 85.7% and 97.6%, respectively. The agreement rate of the two methods was 96.4%, showing a good detection performance. The results indicated that the reMeP72 was of potential value for the serodiagnosis of ASF. Keywords: African swine fever virus; P72 gene; antigenic protein; colloidal gold-based immunochromatographic assay.

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非洲猪瘟多表位蛋白p72 (MeP72)血清诊断的研制与评价。

非洲猪瘟(African swine fever, ASF)是一种严重危害全球养猪业的急性重症传染病。为开发特异性和敏感性高的非洲猪瘟血清诊断试剂，本研究对非洲猪瘟病毒P72蛋白抗原表位进行了分析，构建了与优势线性表位串联的非洲猪瘟多表位融合基因MeP72。在大肠杆菌中制备了重组多表位融合蛋白MeP72 (rep72)。建立了一种基于rep72的胶体金免疫层析法(CGIA)，用于检测ASFV抗体。采用139份猪临床血清样本评估rep72的潜在诊断价值。结果表明，CGIA与猪瘟病毒(CSFV)、猪圆环病毒2型(PCV2)、伪狂犬病毒(PRV)、猪繁殖与呼吸综合征病毒(PRRSV)、猪流感病毒(SIV)等病毒阳性血清无交叉反应，特异性高。敏感性分析显示，CGIA在1:64的稀释倍数下可检出asfv阳性血清。与市售的ASFV检测试剂盒相比，基于reMeP72蛋白的ASFV CGIA检测试剂盒的灵敏度和特异性分别为85.7%和97.6%。两种方法的符合率为96.4%，具有良好的检测性能。结果表明，rep72在ASF的血清诊断中具有潜在的应用价值。关键词:非洲猪瘟病毒;P72基因;抗原蛋白;胶体金免疫层析法。

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来源期刊

Acta virologica 医学-病毒学

CiteScore

3.10

自引率

11.80%

发文量

审稿时长

>12 weeks

期刊介绍： Acta virologica is an international journal of predominantly molecular and cellular virology. Acta virologica aims to publish papers reporting original results of fundamental and applied research mainly on human, animal and plant viruses at cellular and molecular level. As a matter of tradition, also rickettsiae are included. Areas of interest are virus structure and morphology, molecular biology of virus-cell interactions, molecular genetics of viruses, pathogenesis of viral diseases, viral immunology, vaccines, antiviral drugs and viral diagnostics.