文献类型： 外文期刊

作者： He, Wan-qin ¹ ; Wu, Jia-yu ² ; Ren, Yi-yi ² ; Zhou, Xue-ping ¹ ; Zhang, Song-bai ⁴ ; Qian, Ya-juan ¹ ; Li, Fang-fang ³ ;

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

2.Zhejiang Univ, Dept Appl Biol Sci, Coll Agr & Biotechnol, Hangzhou 310058, Peoples R China

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

4.Chinese Acad Agr Sci, Hunan Plant Protect Inst, Changsha 410125, Peoples R China

关键词： Pepino mosaic virus; Monoclonal antibody; Serological method; Double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA); Dot-ELISA; Tissue print-ELISA; S41-30

期刊名称：JOURNAL OF ZHEJIANG UNIVERSITY-SCIENCE B （ 影响因子：3.066； 五年影响因子：3.057 ）

ISSN： 1673-1581

年卷期： 2020 年 21 卷 10 期

页码：

收录情况： SCI

摘要： Pepino mosaic virus(PepMV) causes severe disease in tomato and other Solanaceous crops around globe. To effectively study and manage this viral disease, researchers need new, sensitive, and high-throughput approaches for viral detection. In this study, we purified PepMV particles from the infectedNicotiana benthamianaplants and used virions to immunize BALB/c mice to prepare hybridomas secreting anti-PepMV monoclonal antibodies (mAbs). A panel of highly specific and sensitive murine mAbs (15B2, 8H6, 23D11, 20D9, 3A6, and 8E3) could be produced through cell fusion, antibody selection, and cell cloning. Using the mAbs as the detection antibodies, we established double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), Dot-ELISA, and Tissue print-ELISA for detecting PepMV infection in tomato plants. Resulting data on sensitivity analysis assays showed that both DAS-ELISA and Dot-ELISA can efficiently monitor the virus in PepMV-infected tissue crude extracts when diluted at 1:1310720 and 1:20480 (weight/volume ratio (w/v), g/mL), respectively. Among the three methods developed, the Tissue print-ELISA was found to be the most practical detection technique. Survey results from field samples by the established serological approaches were verified by reverse transcription polymerase chain reaction (RT-PCR) and DNA sequencing, demonstrating all three serological methods are reliable and effective for monitoring PepMV. Anti-PepMV mAbs and the newly developed DAS-ELISA, Dot-ELISA, and Tissue print-ELISA can benefit PepMV detection and field epidemiological study, and management of this viral disease, which is already widespread in tomato plants in Yunnan Province of China.