CLC number: S41-30
On-line Access: 2020-10-12
Received: 2020-05-17
Revision Accepted: 2020-07-30
Crosschecked: 2020-09-08
Cited: 0
Clicked: 3915
Citations: Bibtex RefMan EndNote GB/T7714
Wan-qin He, Jia-yu Wu, Yi-yi Ren, Xue-ping Zhou, Song-bai Zhang, Ya-juan Qian, Fang-fang Li, Jian-xiang Wu. Highly sensitive serological approaches for Pepino mosaic virus detection[J]. Journal of Zhejiang University Science B, 2020, 21(10): 811-822.
@article{title="Highly sensitive serological approaches for Pepino mosaic virus detection",
author="Wan-qin He, Jia-yu Wu, Yi-yi Ren, Xue-ping Zhou, Song-bai Zhang, Ya-juan Qian, Fang-fang Li, Jian-xiang Wu",
journal="Journal of Zhejiang University Science B",
volume="21",
number="10",
pages="811-822",
year="2020",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B2000255"
}
%0 Journal Article
%T Highly sensitive serological approaches for Pepino mosaic virus detection
%A Wan-qin He
%A Jia-yu Wu
%A Yi-yi Ren
%A Xue-ping Zhou
%A Song-bai Zhang
%A Ya-juan Qian
%A Fang-fang Li
%A Jian-xiang Wu
%J Journal of Zhejiang University SCIENCE B
%V 21
%N 10
%P 811-822
%@ 1673-1581
%D 2020
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B2000255
TY - JOUR
T1 - Highly sensitive serological approaches for Pepino mosaic virus detection
A1 - Wan-qin He
A1 - Jia-yu Wu
A1 - Yi-yi Ren
A1 - Xue-ping Zhou
A1 - Song-bai Zhang
A1 - Ya-juan Qian
A1 - Fang-fang Li
A1 - Jian-xiang Wu
J0 - Journal of Zhejiang University Science B
VL - 21
IS - 10
SP - 811
EP - 822
%@ 1673-1581
Y1 - 2020
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B2000255
Abstract: 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 infected Nicotiana benthamiana plants 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 1:1 310 720 and 1:20 480 (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.
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