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Monoclonal antibody-based serological detection of potato virus M in potato plants and tubers

文献类型： 外文期刊

作者： Zhang Yu 1 ; Gao Yan-ling 2 ; He Wan-qin 1 ; Wang Ya-qin 1 ; Qian Ya-juan 1 ; Zhou Xue-ping 1 ; Wu Jian-xiang 1 ;

作者机构： 1.Zhejiang Univ, Inst Biotechnol, State Key Lab Rice Biol, Hangzhou 310058, Peoples R China

2.Heilongjiang Acad Agr Sci, Potato Res Inst, Harbin 150086, Peoples R China

3.Chinese Acad Agr Sci, State Key Lab Biol Plant Dis & Insect Pests, Inst Plant Protect, Beijing 100193, Peoples R China

关键词： potato virus M; monoclonal antibody; serological assay; antigen-coated plate (ACP)-ELISA; dot-ELISA; Tissue print-ELISA

期刊名称：JOURNAL OF INTEGRATIVE AGRICULTURE （ 影响因子：2.848； 五年影响因子：2.979 ）

ISSN： 2095-3119

年卷期： 2020 年 19 卷 5 期

页码：

收录情况： SCI

摘要： Potato virus M (PVM) is one of the common and economically important potato viruses in potato-growing regions worldwide. To investigate and control this viral disease, efficient and specific detection techniques are needed. In this study, PVM virions were purified from infected potato plants and used as the immunogen to produce hybridomas secreting PVM-specific monoclonal antibodies (MAbs). Four highly specific and sensitive murine MAbs, i.e., 1E1, 2A5, 8A1 and 17G8 were prepared through a conventional hybridoma technology. Using these four MAbs, we have developed an antigen-coated plate (ACP)-ELISA, a dot-ELISA and a Tissue print-ELISAfor detecting PVM infection in potato plants and tubers. PVM could be detected in infected potato plant tissue crude extracts diluted at 1:10240 (w/v, g mL(-1)) by the dot-ELISA or at 1:163840 (w/v, g mL(-1)) by the ACP-ELISA. The Tissue print-ELISA is the quickest and easiest assay among the three established serological assays and is more suitable for onsite large-scale sample detection. Detection results of the field-collected samples showed that PVM is currently widespread in the Yunnan and the Heilongjiang provinces in China. The field sample test results of the developed serological assays were supported by the results from RT-PCR and DNA sequencing. We consider that the newly established ACP-ELISA, dot-ELISA and Tissue print-ELISA can benefit PVM detection in potato plant and tuber samples and field epidemiological studies of PVM. These assays can also facilitate the production of virus-free seed potatoes and breeding for PVM-resistant potato cultivars, leading to the successful prevention of this potato viral disease.

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